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200 000 neonatal foreskin hff1 fibroblasts  (ATCC)


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    ATCC 200 000 neonatal foreskin hff1 fibroblasts
    Figure 1. Generation of human induced pluripotent stem cells (iPSCs) and characterization of iPS2 and iPS4 lines. iPS cells were derived by retroviral transduction of the Yamanaka cocktail (OCT4, KLF4, SOX2, and c-MYC) in neonatal foreskin fibroblasts <t>HFF1.</t> Two iPSC lines (iPS2 and iPS4) were fully characterized. (A): Pictures showing the essential steps of iPSC reprogramming. Upper lane, from left to right: HFF1 fibro- blasts at passage 4 used for the reprogramming experiments, cellular clone 28 days after transduction, AP staining 4 weeks after transduction. Lower lane, from left to right: iPS4 line at passage 10, iPS2 line at passage 8, iPS4 line at passage 12 stained with AP. (B): Immunofluorescence imaging of embryonic stem (ES) cell markers in iPS4 cells. (C): Retroviral silencing in iPS2 and iPS4 determined by quantitative polymerase chain reaction (QPCR). As positive control, HFF1 cells were transduced with a single factor (respectively, HFF1-O, HFF1-K, HFF1-S, and HFF1-M) and harvested 72 hours later for RNA isolation. Immunofluorescence pictures confirming the expression of the transduced factors are reported on the right side. (D): Upper left panel, DNA fingerprinting analysis confirmed the somatic origin of iPS2 and iPS4. Upper right panel, karyotype analysis of the iPS4 line. Lower panel, QPCR analysis of pluripotency-associated genes in H1, H9, iPS2, and iPS4. Results were nor- malized with GAPDH and expressed in comparison to H1.
    200 000 Neonatal Foreskin Hff1 Fibroblasts, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1587 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/200+000+neonatal+foreskin+hff1+fibroblasts/HFF-1/pm20201066-53-5-10
    Average 99 stars, based on 1587 article reviews
    200 000 neonatal foreskin hff1 fibroblasts - by Bioz Stars, 2026-10
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    1) Product Images from "The senescence-related mitochondrial/oxidative stress pathway is repressed in human induced pluripotent stem cells."

    Article Title: The senescence-related mitochondrial/oxidative stress pathway is repressed in human induced pluripotent stem cells.

    Journal: Stem cells (Dayton, Ohio)

    doi: 10.1002/stem.404

    Figure 1. Generation of human induced pluripotent stem cells (iPSCs) and characterization of iPS2 and iPS4 lines. iPS cells were derived by retroviral transduction of the Yamanaka cocktail (OCT4, KLF4, SOX2, and c-MYC) in neonatal foreskin fibroblasts HFF1. Two iPSC lines (iPS2 and iPS4) were fully characterized. (A): Pictures showing the essential steps of iPSC reprogramming. Upper lane, from left to right: HFF1 fibro- blasts at passage 4 used for the reprogramming experiments, cellular clone 28 days after transduction, AP staining 4 weeks after transduction. Lower lane, from left to right: iPS4 line at passage 10, iPS2 line at passage 8, iPS4 line at passage 12 stained with AP. (B): Immunofluorescence imaging of embryonic stem (ES) cell markers in iPS4 cells. (C): Retroviral silencing in iPS2 and iPS4 determined by quantitative polymerase chain reaction (QPCR). As positive control, HFF1 cells were transduced with a single factor (respectively, HFF1-O, HFF1-K, HFF1-S, and HFF1-M) and harvested 72 hours later for RNA isolation. Immunofluorescence pictures confirming the expression of the transduced factors are reported on the right side. (D): Upper left panel, DNA fingerprinting analysis confirmed the somatic origin of iPS2 and iPS4. Upper right panel, karyotype analysis of the iPS4 line. Lower panel, QPCR analysis of pluripotency-associated genes in H1, H9, iPS2, and iPS4. Results were nor- malized with GAPDH and expressed in comparison to H1.
    Figure Legend Snippet: Figure 1. Generation of human induced pluripotent stem cells (iPSCs) and characterization of iPS2 and iPS4 lines. iPS cells were derived by retroviral transduction of the Yamanaka cocktail (OCT4, KLF4, SOX2, and c-MYC) in neonatal foreskin fibroblasts HFF1. Two iPSC lines (iPS2 and iPS4) were fully characterized. (A): Pictures showing the essential steps of iPSC reprogramming. Upper lane, from left to right: HFF1 fibro- blasts at passage 4 used for the reprogramming experiments, cellular clone 28 days after transduction, AP staining 4 weeks after transduction. Lower lane, from left to right: iPS4 line at passage 10, iPS2 line at passage 8, iPS4 line at passage 12 stained with AP. (B): Immunofluorescence imaging of embryonic stem (ES) cell markers in iPS4 cells. (C): Retroviral silencing in iPS2 and iPS4 determined by quantitative polymerase chain reaction (QPCR). As positive control, HFF1 cells were transduced with a single factor (respectively, HFF1-O, HFF1-K, HFF1-S, and HFF1-M) and harvested 72 hours later for RNA isolation. Immunofluorescence pictures confirming the expression of the transduced factors are reported on the right side. (D): Upper left panel, DNA fingerprinting analysis confirmed the somatic origin of iPS2 and iPS4. Upper right panel, karyotype analysis of the iPS4 line. Lower panel, QPCR analysis of pluripotency-associated genes in H1, H9, iPS2, and iPS4. Results were nor- malized with GAPDH and expressed in comparison to H1.

    Techniques Used: Derivative Assay, Retroviral, Transduction, Staining, Imaging, Real-time Polymerase Chain Reaction, Positive Control, Isolation, Expressing, Comparison

    Figure 2. Mitochondrial-related nuclear transcriptional profiling in hESCs and iPSCs. Transcriptome analysis of fibroblasts (HFF1), hESC (H1 and H9), and iPSCs (iPS2 and iPS4) was performed using illumina bead chips. (A): Upper panel, hierarchical clustering. Lower panel, table showing the correlation coefficient (r2) between the samples. Low r2 values (in gray) were detected between fibroblasts and ESCs and between fibroblasts and iPSCs. ES and iPS cells exhibited higher r2 values (in light blue). (B): Venn diagram showing the distinct and overlapping tran- scriptional signatures between the three cell types. Identified signatures are reported. (C): Pathway analysis of the 7,302 genes comprising the housekeeping transcriptional signature revealed several mitochondrial-related pathways. (D): Heatmap figures depicting transcripts related to the oxidative stress response and to mitochondrial biogenesis. Values represent the ratio of the array average signal of the given gene divided by the average signal of HFF1 line (fold change 1.5, p value .01). Downregulated genes are shown in green, and upregulated genes are shown in red. Abbreviations: hESCs, human embryonic stem cells; iPSCs, induced pluripotent stem cells.
    Figure Legend Snippet: Figure 2. Mitochondrial-related nuclear transcriptional profiling in hESCs and iPSCs. Transcriptome analysis of fibroblasts (HFF1), hESC (H1 and H9), and iPSCs (iPS2 and iPS4) was performed using illumina bead chips. (A): Upper panel, hierarchical clustering. Lower panel, table showing the correlation coefficient (r2) between the samples. Low r2 values (in gray) were detected between fibroblasts and ESCs and between fibroblasts and iPSCs. ES and iPS cells exhibited higher r2 values (in light blue). (B): Venn diagram showing the distinct and overlapping tran- scriptional signatures between the three cell types. Identified signatures are reported. (C): Pathway analysis of the 7,302 genes comprising the housekeeping transcriptional signature revealed several mitochondrial-related pathways. (D): Heatmap figures depicting transcripts related to the oxidative stress response and to mitochondrial biogenesis. Values represent the ratio of the array average signal of the given gene divided by the average signal of HFF1 line (fold change 1.5, p value .01). Downregulated genes are shown in green, and upregulated genes are shown in red. Abbreviations: hESCs, human embryonic stem cells; iPSCs, induced pluripotent stem cells.

    Techniques Used:

    Figure 3. Mitochondrial biogenesis and mitochondrial DNA (mtDNA) copy number in undifferentiated and differentiated human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs). To monitor mitochondrial biogenesis in the self-renewal and differentiated states, fibro- blast-like cells were generated from human ESCs and iPSCs. H1- and H9-derived fibroblasts (respectively, H1-DF and H9-DF) and iPS2 and iPS4- derived fibroblasts (respectively, iPS2-DF and iPS4-DF) were obtained as previously described [2, 12]. (A): Quantitative polymerase chain reaction analysis of nuclear factors involved in mitochondrial biogenesis. Expression values were normalized over the expression of actin beta (ACTB) and presented as relative changes compared to HFF1. (B): Quantification of mtDNA copy number within HFF1 and undifferentiated and spontaneously differentiated ES and iPS cells. ***, p .01, two-tailed unpaired student’s t-test, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 versus HFF1. #, p .05, two-tailed unpaired student’s t-test, H1-DF versus H1. ##, p .05, two-tailed unpaired student’s t-test, H9-DF versus H9. ###, p .05, two-tailed unpaired student’s t-test, iPS2-DF versus iPS2. ####, p .05, two-tailed unpaired student’s t-test, iPS4-DF versus iPS4. (C): Mitochondria content was assayed using the mitochondrion-selective dye MitoTracker Green (MTG). Nuclei were counterstained with DAPI and OCT4 immunofluorescence was used to monitor the relationship between mitochondrial mass and self-renewal capacity.
    Figure Legend Snippet: Figure 3. Mitochondrial biogenesis and mitochondrial DNA (mtDNA) copy number in undifferentiated and differentiated human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs). To monitor mitochondrial biogenesis in the self-renewal and differentiated states, fibro- blast-like cells were generated from human ESCs and iPSCs. H1- and H9-derived fibroblasts (respectively, H1-DF and H9-DF) and iPS2 and iPS4- derived fibroblasts (respectively, iPS2-DF and iPS4-DF) were obtained as previously described [2, 12]. (A): Quantitative polymerase chain reaction analysis of nuclear factors involved in mitochondrial biogenesis. Expression values were normalized over the expression of actin beta (ACTB) and presented as relative changes compared to HFF1. (B): Quantification of mtDNA copy number within HFF1 and undifferentiated and spontaneously differentiated ES and iPS cells. ***, p .01, two-tailed unpaired student’s t-test, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 versus HFF1. #, p .05, two-tailed unpaired student’s t-test, H1-DF versus H1. ##, p .05, two-tailed unpaired student’s t-test, H9-DF versus H9. ###, p .05, two-tailed unpaired student’s t-test, iPS2-DF versus iPS2. ####, p .05, two-tailed unpaired student’s t-test, iPS4-DF versus iPS4. (C): Mitochondria content was assayed using the mitochondrion-selective dye MitoTracker Green (MTG). Nuclei were counterstained with DAPI and OCT4 immunofluorescence was used to monitor the relationship between mitochondrial mass and self-renewal capacity.

    Techniques Used: Generated, Derivative Assay, Real-time Polymerase Chain Reaction, Expressing, Two Tailed Test

    Figure 4. Mitochondrial morphology and distribution within hESCs and iPSCs. Transmission electron microscopy (TEM) was used to investi- gate the morphological features of the mitochondrial network within differentiated and undifferentiated cells. (A): TEM pictures at low and high magnification of HFF1 somatic fibroblasts, undifferentiated H1 and iPS4 lines, and their derived fibroblast-like cells, H1-DF and iPS4-DF, respec- tively. Asterisks show mitochondria with tubular elongated morphology; arrowheads indicate round-shaped mitochondria with poorly developed cristae. This latter feature was observed as the most common one in undifferentiated ES and iPS cells. (B): Graphs reporting the measurement of mitochondrial diameters. D1 and D2 represent, respectively, the major and minor axes, as shown in the scheme on the left. D1 diameter appeared significantly lower in undifferentiated ES and iPS cells compared to HFF1, whereas it was increased in derived-fibroblasts in comparison to their respective undifferentiated cells. An inverse pattern could be observed for the D2 diameter. ***, p .001, two-tailed unpaired student’s t-test, hESCs versus HFF1 and iPSCs versus HFF1. #, p .01, two-tailed unpaired student’s t-test, hESC-DF versus hESCs. ##, p .05, two-tailed unpaired student’s t-test, iPSC-DF versus iPSCs. Fifty mitochondria were counted for each cell type. Bars represent standard deviation. (C): TEM overview images showing mitochondria localization within undifferentiated H1 and iPS4 cell lines. Arrows indicate round-shaped mito- chondria clustering at the two sides of the nuclei. Abbreviations: DF, fibroblast-like differentiated cells; ESCs, embryonic stem cells; iPSCs, induced pluripotent stem cells.
    Figure Legend Snippet: Figure 4. Mitochondrial morphology and distribution within hESCs and iPSCs. Transmission electron microscopy (TEM) was used to investi- gate the morphological features of the mitochondrial network within differentiated and undifferentiated cells. (A): TEM pictures at low and high magnification of HFF1 somatic fibroblasts, undifferentiated H1 and iPS4 lines, and their derived fibroblast-like cells, H1-DF and iPS4-DF, respec- tively. Asterisks show mitochondria with tubular elongated morphology; arrowheads indicate round-shaped mitochondria with poorly developed cristae. This latter feature was observed as the most common one in undifferentiated ES and iPS cells. (B): Graphs reporting the measurement of mitochondrial diameters. D1 and D2 represent, respectively, the major and minor axes, as shown in the scheme on the left. D1 diameter appeared significantly lower in undifferentiated ES and iPS cells compared to HFF1, whereas it was increased in derived-fibroblasts in comparison to their respective undifferentiated cells. An inverse pattern could be observed for the D2 diameter. ***, p .001, two-tailed unpaired student’s t-test, hESCs versus HFF1 and iPSCs versus HFF1. #, p .01, two-tailed unpaired student’s t-test, hESC-DF versus hESCs. ##, p .05, two-tailed unpaired student’s t-test, iPSC-DF versus iPSCs. Fifty mitochondria were counted for each cell type. Bars represent standard deviation. (C): TEM overview images showing mitochondria localization within undifferentiated H1 and iPS4 cell lines. Arrows indicate round-shaped mito- chondria clustering at the two sides of the nuclei. Abbreviations: DF, fibroblast-like differentiated cells; ESCs, embryonic stem cells; iPSCs, induced pluripotent stem cells.

    Techniques Used: Transmission Assay, Electron Microscopy, Derivative Assay, Comparison, Two Tailed Test, Standard Deviation

    Figure 5. Aerobic and anaerobic energy gen- eration in human embryonic stem cells (hESCs) and iPSCs. Cellular energy can be obtained either by aerobic respiration, which produces high amount of ATP and requires ox- ygen and active mitochondria, or by anaerobic respiration followed by lactic acid formation. To test energy generation in undifferentiated and differentiated iPS cells, ATP content and lactate generation were measured. (A): Cellular ATP level in HFF1 somatic fibroblasts, undif- ferentiated hESC H1 and H9 lines, undifferen- tiated iPSC iPS2 and iPS4 lines, and derived fibroblasts H1-DF, H9-DF, iPS2-DF, and iPS4- DF cells. ***, p .01, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 ver- sus HFF1. #, p .05, H1-DF versus H1. ##, p .05, H9-DF versus H9. ###, p .05, iPS2- DF versus iPS2. ####, p .05, iPS4-DF ver- sus iPS4. (B): Levels of extracellular lactate in undifferentiated and differentiated cells in comparison to HFF1. ***, p .01, H1 versus HFF1, and H9 versus HFF1. **, p .05, iPS2 versus HFF1 and iPS4 versus HFF1. #, p .01, H1-DF versus H1. ##, p .01, H9-DF versus H9. ###, p .05, iPS2-DF versus iPS2. ####, p .05, iPS4-DF versus iPS4. Abbrevi- ations: DF, fibroblast-like differentiated cells; iPS, induced pluripotent stem.
    Figure Legend Snippet: Figure 5. Aerobic and anaerobic energy gen- eration in human embryonic stem cells (hESCs) and iPSCs. Cellular energy can be obtained either by aerobic respiration, which produces high amount of ATP and requires ox- ygen and active mitochondria, or by anaerobic respiration followed by lactic acid formation. To test energy generation in undifferentiated and differentiated iPS cells, ATP content and lactate generation were measured. (A): Cellular ATP level in HFF1 somatic fibroblasts, undif- ferentiated hESC H1 and H9 lines, undifferen- tiated iPSC iPS2 and iPS4 lines, and derived fibroblasts H1-DF, H9-DF, iPS2-DF, and iPS4- DF cells. ***, p .01, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 ver- sus HFF1. #, p .05, H1-DF versus H1. ##, p .05, H9-DF versus H9. ###, p .05, iPS2- DF versus iPS2. ####, p .05, iPS4-DF ver- sus iPS4. (B): Levels of extracellular lactate in undifferentiated and differentiated cells in comparison to HFF1. ***, p .01, H1 versus HFF1, and H9 versus HFF1. **, p .05, iPS2 versus HFF1 and iPS4 versus HFF1. #, p .01, H1-DF versus H1. ##, p .01, H9-DF versus H9. ###, p .05, iPS2-DF versus iPS2. ####, p .05, iPS4-DF versus iPS4. Abbrevi- ations: DF, fibroblast-like differentiated cells; iPS, induced pluripotent stem.

    Techniques Used: Derivative Assay, Comparison

    Figure 6. Oxidative-mediated damage in human embryonic stem cells (hESCs) and iPSCs. Highly active mitochondria may generate reactive oxygen species (ROS) as byproducts. ROS-related damage to proteins, DNA, and lipids was evaluated in undifferentiated and differentiated cells. (A): Left panel, representative image for OxyBlot assay, which detects the level of oxidatively modified proteins. Membranes were stripped and reprobed with GAPDH as a loading control. Right panel, average densitometry analysis normalized over GAPDH expression and presented as area under the curve (AUC) values. **, p .05, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 versus HFF1. #, p .05, H1- DF versus H1. ##, p .05, H9-DF versus H9. ###, p .05, iPS2-DF versus iPS2. ####, p .05, iPS4-DF versus iPS4. (B): Immunofluores- cence staining with 8-OHdG of untreated cells and cells exposed to 500 uM of H202 for 2 hours. Nuclei were counterstained with DAPI. (C): Levels of oxidatively modified lipids were monitored by measuring lipid hydroperoxidases (LPO), as previously shown [37]. ***, p .01, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 versus HFF1. #, p .05, H1-DF versus H1. ##, p .05, H9-DF versus H9. ###, p .05, iPS2-DF versus iPS2. ####, p .05, iPS4-DF versus iPS4. (D): Upper panel, representative immunoblot image of glutathione peroxidase 1 (GPX1), the most abundant glutathione peroxidase isozyme in mammalian cells. Membranes were stripped and reprobed with GAPDH as a loading control. Lower panel, average densitometry analysis normalized over GAPDH expression and presented as AUC values. ***, p .01, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 versus HFF1. ###, p .01, iPS2-DF versus iPS2. ####, p .01, iPS4-DF versus iPS4. Abbreviations: DF, fibroblast-like differentiated cells; iPS, induced pluripotent stem.
    Figure Legend Snippet: Figure 6. Oxidative-mediated damage in human embryonic stem cells (hESCs) and iPSCs. Highly active mitochondria may generate reactive oxygen species (ROS) as byproducts. ROS-related damage to proteins, DNA, and lipids was evaluated in undifferentiated and differentiated cells. (A): Left panel, representative image for OxyBlot assay, which detects the level of oxidatively modified proteins. Membranes were stripped and reprobed with GAPDH as a loading control. Right panel, average densitometry analysis normalized over GAPDH expression and presented as area under the curve (AUC) values. **, p .05, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 versus HFF1. #, p .05, H1- DF versus H1. ##, p .05, H9-DF versus H9. ###, p .05, iPS2-DF versus iPS2. ####, p .05, iPS4-DF versus iPS4. (B): Immunofluores- cence staining with 8-OHdG of untreated cells and cells exposed to 500 uM of H202 for 2 hours. Nuclei were counterstained with DAPI. (C): Levels of oxidatively modified lipids were monitored by measuring lipid hydroperoxidases (LPO), as previously shown [37]. ***, p .01, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 versus HFF1. #, p .05, H1-DF versus H1. ##, p .05, H9-DF versus H9. ###, p .05, iPS2-DF versus iPS2. ####, p .05, iPS4-DF versus iPS4. (D): Upper panel, representative immunoblot image of glutathione peroxidase 1 (GPX1), the most abundant glutathione peroxidase isozyme in mammalian cells. Membranes were stripped and reprobed with GAPDH as a loading control. Lower panel, average densitometry analysis normalized over GAPDH expression and presented as AUC values. ***, p .01, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 versus HFF1. ###, p .01, iPS2-DF versus iPS2. ####, p .01, iPS4-DF versus iPS4. Abbreviations: DF, fibroblast-like differentiated cells; iPS, induced pluripotent stem.

    Techniques Used: Control, Expressing, Staining, Western Blot

    Related Articles

    Transduction:

    Article Title: The senescence-related mitochondrial/oxidative stress pathway is repressed in human induced pluripotent stem cells.
    Article Snippet: After 30 hours, medium was collected, centrifuged at 20,000 rpm for 2 hours, filtered, and supplemented with 4 lg/ml of Polybrene (Sigma, St. Louis, http://www.sigmaaldrich.com). .. For generation of human iPSCs, 200.000 neonatal foreskin HFF1 fibroblasts (ATCC, #ATCCSCRC-1041, http://www.atcc.org) were seeded and transduced with the retroviral cocktail on days 1 and 2. ..

    Retroviral:

    Article Title: The senescence-related mitochondrial/oxidative stress pathway is repressed in human induced pluripotent stem cells.
    Article Snippet: After 30 hours, medium was collected, centrifuged at 20,000 rpm for 2 hours, filtered, and supplemented with 4 lg/ml of Polybrene (Sigma, St. Louis, http://www.sigmaaldrich.com). .. For generation of human iPSCs, 200.000 neonatal foreskin HFF1 fibroblasts (ATCC, #ATCCSCRC-1041, http://www.atcc.org) were seeded and transduced with the retroviral cocktail on days 1 and 2. ..



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    200 000 neonatal foreskin hff1 fibroblasts  (ATCC)
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    ATCC 200 000 neonatal foreskin hff1 fibroblasts
    Figure 1. Generation of human induced pluripotent stem cells (iPSCs) and characterization of iPS2 and iPS4 lines. iPS cells were derived by retroviral transduction of the Yamanaka cocktail (OCT4, KLF4, SOX2, and c-MYC) in neonatal foreskin fibroblasts <t>HFF1.</t> Two iPSC lines (iPS2 and iPS4) were fully characterized. (A): Pictures showing the essential steps of iPSC reprogramming. Upper lane, from left to right: HFF1 fibro- blasts at passage 4 used for the reprogramming experiments, cellular clone 28 days after transduction, AP staining 4 weeks after transduction. Lower lane, from left to right: iPS4 line at passage 10, iPS2 line at passage 8, iPS4 line at passage 12 stained with AP. (B): Immunofluorescence imaging of embryonic stem (ES) cell markers in iPS4 cells. (C): Retroviral silencing in iPS2 and iPS4 determined by quantitative polymerase chain reaction (QPCR). As positive control, HFF1 cells were transduced with a single factor (respectively, HFF1-O, HFF1-K, HFF1-S, and HFF1-M) and harvested 72 hours later for RNA isolation. Immunofluorescence pictures confirming the expression of the transduced factors are reported on the right side. (D): Upper left panel, DNA fingerprinting analysis confirmed the somatic origin of iPS2 and iPS4. Upper right panel, karyotype analysis of the iPS4 line. Lower panel, QPCR analysis of pluripotency-associated genes in H1, H9, iPS2, and iPS4. Results were nor- malized with GAPDH and expressed in comparison to H1.
    200 000 Neonatal Foreskin Hff1 Fibroblasts, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/200+000+neonatal+foreskin+hff1+fibroblasts/HFF-1/pm20201066-53-5-10
    Average 99 stars, based on 1 article reviews
    200 000 neonatal foreskin hff1 fibroblasts - by Bioz Stars, 2026-10
    99/100 stars
    		  Buy from Supplier

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    Figure 1. Generation of human induced pluripotent stem cells (iPSCs) and characterization of iPS2 and iPS4 lines. iPS cells were derived by retroviral transduction of the Yamanaka cocktail (OCT4, KLF4, SOX2, and c-MYC) in neonatal foreskin fibroblasts HFF1. Two iPSC lines (iPS2 and iPS4) were fully characterized. (A): Pictures showing the essential steps of iPSC reprogramming. Upper lane, from left to right: HFF1 fibro- blasts at passage 4 used for the reprogramming experiments, cellular clone 28 days after transduction, AP staining 4 weeks after transduction. Lower lane, from left to right: iPS4 line at passage 10, iPS2 line at passage 8, iPS4 line at passage 12 stained with AP. (B): Immunofluorescence imaging of embryonic stem (ES) cell markers in iPS4 cells. (C): Retroviral silencing in iPS2 and iPS4 determined by quantitative polymerase chain reaction (QPCR). As positive control, HFF1 cells were transduced with a single factor (respectively, HFF1-O, HFF1-K, HFF1-S, and HFF1-M) and harvested 72 hours later for RNA isolation. Immunofluorescence pictures confirming the expression of the transduced factors are reported on the right side. (D): Upper left panel, DNA fingerprinting analysis confirmed the somatic origin of iPS2 and iPS4. Upper right panel, karyotype analysis of the iPS4 line. Lower panel, QPCR analysis of pluripotency-associated genes in H1, H9, iPS2, and iPS4. Results were nor- malized with GAPDH and expressed in comparison to H1.

    Journal: Stem cells (Dayton, Ohio)

    Article Title: The senescence-related mitochondrial/oxidative stress pathway is repressed in human induced pluripotent stem cells.

    doi: 10.1002/stem.404

    Figure Lengend Snippet: Figure 1. Generation of human induced pluripotent stem cells (iPSCs) and characterization of iPS2 and iPS4 lines. iPS cells were derived by retroviral transduction of the Yamanaka cocktail (OCT4, KLF4, SOX2, and c-MYC) in neonatal foreskin fibroblasts HFF1. Two iPSC lines (iPS2 and iPS4) were fully characterized. (A): Pictures showing the essential steps of iPSC reprogramming. Upper lane, from left to right: HFF1 fibro- blasts at passage 4 used for the reprogramming experiments, cellular clone 28 days after transduction, AP staining 4 weeks after transduction. Lower lane, from left to right: iPS4 line at passage 10, iPS2 line at passage 8, iPS4 line at passage 12 stained with AP. (B): Immunofluorescence imaging of embryonic stem (ES) cell markers in iPS4 cells. (C): Retroviral silencing in iPS2 and iPS4 determined by quantitative polymerase chain reaction (QPCR). As positive control, HFF1 cells were transduced with a single factor (respectively, HFF1-O, HFF1-K, HFF1-S, and HFF1-M) and harvested 72 hours later for RNA isolation. Immunofluorescence pictures confirming the expression of the transduced factors are reported on the right side. (D): Upper left panel, DNA fingerprinting analysis confirmed the somatic origin of iPS2 and iPS4. Upper right panel, karyotype analysis of the iPS4 line. Lower panel, QPCR analysis of pluripotency-associated genes in H1, H9, iPS2, and iPS4. Results were nor- malized with GAPDH and expressed in comparison to H1.

    Article Snippet: For generation of human iPSCs, 200.000 neonatal foreskin HFF1 fibroblasts (ATCC, #ATCCSCRC-1041, http://www.atcc.org) were seeded and transduced with the retroviral cocktail on days 1 and 2.

    Techniques: Derivative Assay, Retroviral, Transduction, Staining, Imaging, Real-time Polymerase Chain Reaction, Positive Control, Isolation, Expressing, Comparison

    Figure 2. Mitochondrial-related nuclear transcriptional profiling in hESCs and iPSCs. Transcriptome analysis of fibroblasts (HFF1), hESC (H1 and H9), and iPSCs (iPS2 and iPS4) was performed using illumina bead chips. (A): Upper panel, hierarchical clustering. Lower panel, table showing the correlation coefficient (r2) between the samples. Low r2 values (in gray) were detected between fibroblasts and ESCs and between fibroblasts and iPSCs. ES and iPS cells exhibited higher r2 values (in light blue). (B): Venn diagram showing the distinct and overlapping tran- scriptional signatures between the three cell types. Identified signatures are reported. (C): Pathway analysis of the 7,302 genes comprising the housekeeping transcriptional signature revealed several mitochondrial-related pathways. (D): Heatmap figures depicting transcripts related to the oxidative stress response and to mitochondrial biogenesis. Values represent the ratio of the array average signal of the given gene divided by the average signal of HFF1 line (fold change 1.5, p value .01). Downregulated genes are shown in green, and upregulated genes are shown in red. Abbreviations: hESCs, human embryonic stem cells; iPSCs, induced pluripotent stem cells.

    Journal: Stem cells (Dayton, Ohio)

    Article Title: The senescence-related mitochondrial/oxidative stress pathway is repressed in human induced pluripotent stem cells.

    doi: 10.1002/stem.404

    Figure Lengend Snippet: Figure 2. Mitochondrial-related nuclear transcriptional profiling in hESCs and iPSCs. Transcriptome analysis of fibroblasts (HFF1), hESC (H1 and H9), and iPSCs (iPS2 and iPS4) was performed using illumina bead chips. (A): Upper panel, hierarchical clustering. Lower panel, table showing the correlation coefficient (r2) between the samples. Low r2 values (in gray) were detected between fibroblasts and ESCs and between fibroblasts and iPSCs. ES and iPS cells exhibited higher r2 values (in light blue). (B): Venn diagram showing the distinct and overlapping tran- scriptional signatures between the three cell types. Identified signatures are reported. (C): Pathway analysis of the 7,302 genes comprising the housekeeping transcriptional signature revealed several mitochondrial-related pathways. (D): Heatmap figures depicting transcripts related to the oxidative stress response and to mitochondrial biogenesis. Values represent the ratio of the array average signal of the given gene divided by the average signal of HFF1 line (fold change 1.5, p value .01). Downregulated genes are shown in green, and upregulated genes are shown in red. Abbreviations: hESCs, human embryonic stem cells; iPSCs, induced pluripotent stem cells.

    Article Snippet: For generation of human iPSCs, 200.000 neonatal foreskin HFF1 fibroblasts (ATCC, #ATCCSCRC-1041, http://www.atcc.org) were seeded and transduced with the retroviral cocktail on days 1 and 2.

    Techniques:

    Figure 3. Mitochondrial biogenesis and mitochondrial DNA (mtDNA) copy number in undifferentiated and differentiated human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs). To monitor mitochondrial biogenesis in the self-renewal and differentiated states, fibro- blast-like cells were generated from human ESCs and iPSCs. H1- and H9-derived fibroblasts (respectively, H1-DF and H9-DF) and iPS2 and iPS4- derived fibroblasts (respectively, iPS2-DF and iPS4-DF) were obtained as previously described [2, 12]. (A): Quantitative polymerase chain reaction analysis of nuclear factors involved in mitochondrial biogenesis. Expression values were normalized over the expression of actin beta (ACTB) and presented as relative changes compared to HFF1. (B): Quantification of mtDNA copy number within HFF1 and undifferentiated and spontaneously differentiated ES and iPS cells. ***, p .01, two-tailed unpaired student’s t-test, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 versus HFF1. #, p .05, two-tailed unpaired student’s t-test, H1-DF versus H1. ##, p .05, two-tailed unpaired student’s t-test, H9-DF versus H9. ###, p .05, two-tailed unpaired student’s t-test, iPS2-DF versus iPS2. ####, p .05, two-tailed unpaired student’s t-test, iPS4-DF versus iPS4. (C): Mitochondria content was assayed using the mitochondrion-selective dye MitoTracker Green (MTG). Nuclei were counterstained with DAPI and OCT4 immunofluorescence was used to monitor the relationship between mitochondrial mass and self-renewal capacity.

    Journal: Stem cells (Dayton, Ohio)

    Article Title: The senescence-related mitochondrial/oxidative stress pathway is repressed in human induced pluripotent stem cells.

    doi: 10.1002/stem.404

    Figure Lengend Snippet: Figure 3. Mitochondrial biogenesis and mitochondrial DNA (mtDNA) copy number in undifferentiated and differentiated human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs). To monitor mitochondrial biogenesis in the self-renewal and differentiated states, fibro- blast-like cells were generated from human ESCs and iPSCs. H1- and H9-derived fibroblasts (respectively, H1-DF and H9-DF) and iPS2 and iPS4- derived fibroblasts (respectively, iPS2-DF and iPS4-DF) were obtained as previously described [2, 12]. (A): Quantitative polymerase chain reaction analysis of nuclear factors involved in mitochondrial biogenesis. Expression values were normalized over the expression of actin beta (ACTB) and presented as relative changes compared to HFF1. (B): Quantification of mtDNA copy number within HFF1 and undifferentiated and spontaneously differentiated ES and iPS cells. ***, p .01, two-tailed unpaired student’s t-test, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 versus HFF1. #, p .05, two-tailed unpaired student’s t-test, H1-DF versus H1. ##, p .05, two-tailed unpaired student’s t-test, H9-DF versus H9. ###, p .05, two-tailed unpaired student’s t-test, iPS2-DF versus iPS2. ####, p .05, two-tailed unpaired student’s t-test, iPS4-DF versus iPS4. (C): Mitochondria content was assayed using the mitochondrion-selective dye MitoTracker Green (MTG). Nuclei were counterstained with DAPI and OCT4 immunofluorescence was used to monitor the relationship between mitochondrial mass and self-renewal capacity.

    Article Snippet: For generation of human iPSCs, 200.000 neonatal foreskin HFF1 fibroblasts (ATCC, #ATCCSCRC-1041, http://www.atcc.org) were seeded and transduced with the retroviral cocktail on days 1 and 2.

    Techniques: Generated, Derivative Assay, Real-time Polymerase Chain Reaction, Expressing, Two Tailed Test

    Figure 4. Mitochondrial morphology and distribution within hESCs and iPSCs. Transmission electron microscopy (TEM) was used to investi- gate the morphological features of the mitochondrial network within differentiated and undifferentiated cells. (A): TEM pictures at low and high magnification of HFF1 somatic fibroblasts, undifferentiated H1 and iPS4 lines, and their derived fibroblast-like cells, H1-DF and iPS4-DF, respec- tively. Asterisks show mitochondria with tubular elongated morphology; arrowheads indicate round-shaped mitochondria with poorly developed cristae. This latter feature was observed as the most common one in undifferentiated ES and iPS cells. (B): Graphs reporting the measurement of mitochondrial diameters. D1 and D2 represent, respectively, the major and minor axes, as shown in the scheme on the left. D1 diameter appeared significantly lower in undifferentiated ES and iPS cells compared to HFF1, whereas it was increased in derived-fibroblasts in comparison to their respective undifferentiated cells. An inverse pattern could be observed for the D2 diameter. ***, p .001, two-tailed unpaired student’s t-test, hESCs versus HFF1 and iPSCs versus HFF1. #, p .01, two-tailed unpaired student’s t-test, hESC-DF versus hESCs. ##, p .05, two-tailed unpaired student’s t-test, iPSC-DF versus iPSCs. Fifty mitochondria were counted for each cell type. Bars represent standard deviation. (C): TEM overview images showing mitochondria localization within undifferentiated H1 and iPS4 cell lines. Arrows indicate round-shaped mito- chondria clustering at the two sides of the nuclei. Abbreviations: DF, fibroblast-like differentiated cells; ESCs, embryonic stem cells; iPSCs, induced pluripotent stem cells.

    Journal: Stem cells (Dayton, Ohio)

    Article Title: The senescence-related mitochondrial/oxidative stress pathway is repressed in human induced pluripotent stem cells.

    doi: 10.1002/stem.404

    Figure Lengend Snippet: Figure 4. Mitochondrial morphology and distribution within hESCs and iPSCs. Transmission electron microscopy (TEM) was used to investi- gate the morphological features of the mitochondrial network within differentiated and undifferentiated cells. (A): TEM pictures at low and high magnification of HFF1 somatic fibroblasts, undifferentiated H1 and iPS4 lines, and their derived fibroblast-like cells, H1-DF and iPS4-DF, respec- tively. Asterisks show mitochondria with tubular elongated morphology; arrowheads indicate round-shaped mitochondria with poorly developed cristae. This latter feature was observed as the most common one in undifferentiated ES and iPS cells. (B): Graphs reporting the measurement of mitochondrial diameters. D1 and D2 represent, respectively, the major and minor axes, as shown in the scheme on the left. D1 diameter appeared significantly lower in undifferentiated ES and iPS cells compared to HFF1, whereas it was increased in derived-fibroblasts in comparison to their respective undifferentiated cells. An inverse pattern could be observed for the D2 diameter. ***, p .001, two-tailed unpaired student’s t-test, hESCs versus HFF1 and iPSCs versus HFF1. #, p .01, two-tailed unpaired student’s t-test, hESC-DF versus hESCs. ##, p .05, two-tailed unpaired student’s t-test, iPSC-DF versus iPSCs. Fifty mitochondria were counted for each cell type. Bars represent standard deviation. (C): TEM overview images showing mitochondria localization within undifferentiated H1 and iPS4 cell lines. Arrows indicate round-shaped mito- chondria clustering at the two sides of the nuclei. Abbreviations: DF, fibroblast-like differentiated cells; ESCs, embryonic stem cells; iPSCs, induced pluripotent stem cells.

    Article Snippet: For generation of human iPSCs, 200.000 neonatal foreskin HFF1 fibroblasts (ATCC, #ATCCSCRC-1041, http://www.atcc.org) were seeded and transduced with the retroviral cocktail on days 1 and 2.

    Techniques: Transmission Assay, Electron Microscopy, Derivative Assay, Comparison, Two Tailed Test, Standard Deviation

    Figure 5. Aerobic and anaerobic energy gen- eration in human embryonic stem cells (hESCs) and iPSCs. Cellular energy can be obtained either by aerobic respiration, which produces high amount of ATP and requires ox- ygen and active mitochondria, or by anaerobic respiration followed by lactic acid formation. To test energy generation in undifferentiated and differentiated iPS cells, ATP content and lactate generation were measured. (A): Cellular ATP level in HFF1 somatic fibroblasts, undif- ferentiated hESC H1 and H9 lines, undifferen- tiated iPSC iPS2 and iPS4 lines, and derived fibroblasts H1-DF, H9-DF, iPS2-DF, and iPS4- DF cells. ***, p .01, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 ver- sus HFF1. #, p .05, H1-DF versus H1. ##, p .05, H9-DF versus H9. ###, p .05, iPS2- DF versus iPS2. ####, p .05, iPS4-DF ver- sus iPS4. (B): Levels of extracellular lactate in undifferentiated and differentiated cells in comparison to HFF1. ***, p .01, H1 versus HFF1, and H9 versus HFF1. **, p .05, iPS2 versus HFF1 and iPS4 versus HFF1. #, p .01, H1-DF versus H1. ##, p .01, H9-DF versus H9. ###, p .05, iPS2-DF versus iPS2. ####, p .05, iPS4-DF versus iPS4. Abbrevi- ations: DF, fibroblast-like differentiated cells; iPS, induced pluripotent stem.

    Journal: Stem cells (Dayton, Ohio)

    Article Title: The senescence-related mitochondrial/oxidative stress pathway is repressed in human induced pluripotent stem cells.

    doi: 10.1002/stem.404

    Figure Lengend Snippet: Figure 5. Aerobic and anaerobic energy gen- eration in human embryonic stem cells (hESCs) and iPSCs. Cellular energy can be obtained either by aerobic respiration, which produces high amount of ATP and requires ox- ygen and active mitochondria, or by anaerobic respiration followed by lactic acid formation. To test energy generation in undifferentiated and differentiated iPS cells, ATP content and lactate generation were measured. (A): Cellular ATP level in HFF1 somatic fibroblasts, undif- ferentiated hESC H1 and H9 lines, undifferen- tiated iPSC iPS2 and iPS4 lines, and derived fibroblasts H1-DF, H9-DF, iPS2-DF, and iPS4- DF cells. ***, p .01, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 ver- sus HFF1. #, p .05, H1-DF versus H1. ##, p .05, H9-DF versus H9. ###, p .05, iPS2- DF versus iPS2. ####, p .05, iPS4-DF ver- sus iPS4. (B): Levels of extracellular lactate in undifferentiated and differentiated cells in comparison to HFF1. ***, p .01, H1 versus HFF1, and H9 versus HFF1. **, p .05, iPS2 versus HFF1 and iPS4 versus HFF1. #, p .01, H1-DF versus H1. ##, p .01, H9-DF versus H9. ###, p .05, iPS2-DF versus iPS2. ####, p .05, iPS4-DF versus iPS4. Abbrevi- ations: DF, fibroblast-like differentiated cells; iPS, induced pluripotent stem.

    Article Snippet: For generation of human iPSCs, 200.000 neonatal foreskin HFF1 fibroblasts (ATCC, #ATCCSCRC-1041, http://www.atcc.org) were seeded and transduced with the retroviral cocktail on days 1 and 2.

    Techniques: Derivative Assay, Comparison

    Figure 6. Oxidative-mediated damage in human embryonic stem cells (hESCs) and iPSCs. Highly active mitochondria may generate reactive oxygen species (ROS) as byproducts. ROS-related damage to proteins, DNA, and lipids was evaluated in undifferentiated and differentiated cells. (A): Left panel, representative image for OxyBlot assay, which detects the level of oxidatively modified proteins. Membranes were stripped and reprobed with GAPDH as a loading control. Right panel, average densitometry analysis normalized over GAPDH expression and presented as area under the curve (AUC) values. **, p .05, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 versus HFF1. #, p .05, H1- DF versus H1. ##, p .05, H9-DF versus H9. ###, p .05, iPS2-DF versus iPS2. ####, p .05, iPS4-DF versus iPS4. (B): Immunofluores- cence staining with 8-OHdG of untreated cells and cells exposed to 500 uM of H202 for 2 hours. Nuclei were counterstained with DAPI. (C): Levels of oxidatively modified lipids were monitored by measuring lipid hydroperoxidases (LPO), as previously shown [37]. ***, p .01, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 versus HFF1. #, p .05, H1-DF versus H1. ##, p .05, H9-DF versus H9. ###, p .05, iPS2-DF versus iPS2. ####, p .05, iPS4-DF versus iPS4. (D): Upper panel, representative immunoblot image of glutathione peroxidase 1 (GPX1), the most abundant glutathione peroxidase isozyme in mammalian cells. Membranes were stripped and reprobed with GAPDH as a loading control. Lower panel, average densitometry analysis normalized over GAPDH expression and presented as AUC values. ***, p .01, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 versus HFF1. ###, p .01, iPS2-DF versus iPS2. ####, p .01, iPS4-DF versus iPS4. Abbreviations: DF, fibroblast-like differentiated cells; iPS, induced pluripotent stem.

    Journal: Stem cells (Dayton, Ohio)

    Article Title: The senescence-related mitochondrial/oxidative stress pathway is repressed in human induced pluripotent stem cells.

    doi: 10.1002/stem.404

    Figure Lengend Snippet: Figure 6. Oxidative-mediated damage in human embryonic stem cells (hESCs) and iPSCs. Highly active mitochondria may generate reactive oxygen species (ROS) as byproducts. ROS-related damage to proteins, DNA, and lipids was evaluated in undifferentiated and differentiated cells. (A): Left panel, representative image for OxyBlot assay, which detects the level of oxidatively modified proteins. Membranes were stripped and reprobed with GAPDH as a loading control. Right panel, average densitometry analysis normalized over GAPDH expression and presented as area under the curve (AUC) values. **, p .05, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 versus HFF1. #, p .05, H1- DF versus H1. ##, p .05, H9-DF versus H9. ###, p .05, iPS2-DF versus iPS2. ####, p .05, iPS4-DF versus iPS4. (B): Immunofluores- cence staining with 8-OHdG of untreated cells and cells exposed to 500 uM of H202 for 2 hours. Nuclei were counterstained with DAPI. (C): Levels of oxidatively modified lipids were monitored by measuring lipid hydroperoxidases (LPO), as previously shown [37]. ***, p .01, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 versus HFF1. #, p .05, H1-DF versus H1. ##, p .05, H9-DF versus H9. ###, p .05, iPS2-DF versus iPS2. ####, p .05, iPS4-DF versus iPS4. (D): Upper panel, representative immunoblot image of glutathione peroxidase 1 (GPX1), the most abundant glutathione peroxidase isozyme in mammalian cells. Membranes were stripped and reprobed with GAPDH as a loading control. Lower panel, average densitometry analysis normalized over GAPDH expression and presented as AUC values. ***, p .01, H1 versus HFF1, H9 versus HFF1, iPS2 versus HFF1, and iPS4 versus HFF1. ###, p .01, iPS2-DF versus iPS2. ####, p .01, iPS4-DF versus iPS4. Abbreviations: DF, fibroblast-like differentiated cells; iPS, induced pluripotent stem.

    Article Snippet: For generation of human iPSCs, 200.000 neonatal foreskin HFF1 fibroblasts (ATCC, #ATCCSCRC-1041, http://www.atcc.org) were seeded and transduced with the retroviral cocktail on days 1 and 2.

    Techniques: Control, Expressing, Staining, Western Blot