Journal: Nanomaterials

Article Title: Ferulic Acid-NLC with Lavandula Essential Oil: A Possible Strategy for Wound-Healing?

doi: 10.3390/nano10050898

Figure Lengend Snippet: Murine fibroblasts viability after contact for 24 h with unloaded and FA-loaded NLC at different concentrations. Results are reported as the mean value ± SD of three separate experiments, each performed in triplicate. * Significance for p < 0.05, comparison between FA-loaded NLC and the respective unloaded NLC.

Article Snippet: The wound-closure properties of unloaded (NLCa and NLCb) and FA-loaded NLC carriers (FA-NLCa and FA-NLCb) were investigated, using murine fibroblasts (CCL-3T3, ATCC, USA).

Techniques: Comparison

Journal: Nanomaterials

Article Title: Ferulic Acid-NLC with Lavandula Essential Oil: A Possible Strategy for Wound-Healing?

doi: 10.3390/nano10050898

Figure Lengend Snippet: Photographs of the gaps among fibroblast cells at 0, 24, and 48 h after incubation with free FA, FA-loaded NLCa, and FA-loaded NLCb at 37 °C in 5% CO 2 and 95% relative humidity.

Article Snippet: The wound-closure properties of unloaded (NLCa and NLCb) and FA-loaded NLC carriers (FA-NLCa and FA-NLCb) were investigated, using murine fibroblasts (CCL-3T3, ATCC, USA).

Techniques: Incubation

Journal: Aging (Albany NY)

Article Title: Naturally occurring mitochondrial-derived peptides are age-dependent regulators of apoptosis, insulin sensitivity, and inflammatory markers

doi: 10.18632/aging.100943

Figure Lengend Snippet: NIT-1 β and 22Rv1 cells were cultured in serum-free (SF) media with 100 nM SHLP or control peptides and assessed for ( A ) cell viability (using the MTS assay) after 72 h; ( B ) apoptosis after 24 h; and ( C ) cell proliferation by BrdU incorporation after 24 h. All data are presented as means ± SEM, and significance was determined by Student's t -tests. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: All cell lines (22RV1, NIT-1 murine β-cells, and 3T3-L1 murine pre-adipocytes) were purchased from ATCC (American tissue culture collection) (Manassas, VA).

Techniques: Cell Culture, Control, MTS Assay, BrdU Incorporation Assay

Journal: Aging (Albany NY)

Article Title: Naturally occurring mitochondrial-derived peptides are age-dependent regulators of apoptosis, insulin sensitivity, and inflammatory markers

doi: 10.18632/aging.100943

Figure Lengend Snippet: The effects of exogenous SHLP2 and SHLP3 on mitochondria were assessed in 22Rv1 cells incubated with 100 nM control peptide, SHLP2, or SHLP3 for 24 h by measuring ( A ) oxygen consumption rate (OCR) performed on a Seahorse XF24 Extracellular Flux Analyzer and ( B ) ATP production. ( C ) Reactive oxygen species (ROS) production as assessed by DHE (dihydroethidium) fluorescence in NIT-1 (top) and 22RV1 (bottom) cells after incubation with 100 nM control peptide, SHLP2, or SHLP3 overnight. All data are presented as means ± SEM. ( D ) NIT-1 β cells were pre-incubated with 100 nM SHLP2 or SHLP3 for 5 h, followed by incubation with 10 μM staurosporine (STS) for 24 h. Apoptosis (pre-G1 peak) was assessed by FACS (fluorescence-activated cell sorting) analysis. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: All cell lines (22RV1, NIT-1 murine β-cells, and 3T3-L1 murine pre-adipocytes) were purchased from ATCC (American tissue culture collection) (Manassas, VA).

Techniques: Incubation, Control, Fluorescence, FACS

Journal: Stem cell research & therapy

Article Title: New characterization and safety evaluation of human limbal stem cells used in clinical application: fidelity of mitotic process and mitotic spindle morphologies.

doi: 10.1186/s13287-023-03586-z

Figure Lengend Snippet: Fig. 2 Evaluation of potency of LSC in 2D cultures. A Positively brown stained nucleus of LSCs for p63 and B Ki67 antibodies specific to human cells shown by the immunocytochemistry (ICC) method. The staining method was conducted at the end of the cultivation period when LSCs dominated the culture and confluence exceeded 90% (magnification 100×). C Viability of LSCs in p0 and p1; D For the Colony-Forming Efficiency (CFE) assay, which was conducted in duplicate for three donors, we assessed the proliferation and differentiation capacity of 1000–1500 seeded limbal stem cells (LSCs) cultured on 3T3 irradiated feeder layers in Petri dishes. After 14 days of culture, a total of 86 ± 13.61 cell colonies were observed, resulting in a calculated CFE of 7.08 ± 1.55%. Data are presented as mean ± SD of all samples. Scale bar 10 mm; E Population doubling time (DT) in hours and F cumulative population doubling (cPDs) were determined for each patient and are represented in histograms

Article Snippet: Briefly, the feeder layer was prepared from a 3T3 murine Swiss albino fibroblast cell line (ATCC® CCL-92TM, VA, USA) preferably for a minimum of two hours and a maximum of 24 h before limbal cell seeding.

Techniques: Staining, Immunocytochemistry, Cell Culture, Irradiation

Journal: Stem cell research & therapy

Article Title: New characterization and safety evaluation of human limbal stem cells used in clinical application: fidelity of mitotic process and mitotic spindle morphologies.

doi: 10.1186/s13287-023-03586-z

Figure Lengend Snippet: Fig. 3 Colony, cell, and nuclear area of LSCs and relation of morphological parameters to p63 levels. A Fluorescence image of live LSC colonies (small cyan nuclei within cells very weakly stained with SiR-actin, magenta) growing within 3T3 feeder fibroblasts (much bigger cyan nuclei with larger nucleoli dots within nucleus, seen as intense cyan dots, within cells that are strongly stained for SiR-actin showing actin filaments, magenta). Cyan, NucBlue-Hoesht-33342; magenta, SiR-actin. Maximal projection of multiple z-stacks. B Area of colonies of LSCs, black dots, mean; black lines, standard deviations. C Nuclear area of individual LSCs within colonies, D Cell area of individual LSCs within colonies, E Nuclear area divided by cell area (nuclear to cytoplasm ratio, N/C) of each individual LSC within colonies. F Immunofluorescence image of fixed LSCs within the colony stained with SiR-actin (actin, magenta) and DAPI (DNA, cyan). Maximal projection of multiple z-stacks. G Nuclear area versus cell area across the population of LSCs within colonies. The black line represents the linear regression fit, or x = y line. H Immunofluorescence image of fixed LSCs within the colony stained with SiR-actin (actin, magenta), DAPI (DNA, cyan), and secondary antibody AlexaFluor488 conjugated to primary antibody raised against p63 4A4 (yellow). Maximal projection of multiple z-stacks. (left), p63 intensity in each cell versus its total area (middle), or its nuclear area (right). Black lines in (H) represent linear regression fit. I Immunofluorescence image of LSCs within the colony stained with secondary antibody AF647 conjugated to primary antibody raised against p63-α (dark blue), secondary antibody AF488 conjugated to primary antibody raised against p63 4A4 (magenta), DAPI (DNA, gray), and secondary antibody AF594 conjugated to primary antibody raised against tubulin (green). Maximal projection of multiple z-stacks. (left) p63-α versus p63 4A4 signal intensity in each cell; p63-α (middle) and p63 4A4 (right) intensity versus their nuclear area. Black lines represent linear regression fit

Article Snippet: Briefly, the feeder layer was prepared from a 3T3 murine Swiss albino fibroblast cell line (ATCC® CCL-92TM, VA, USA) preferably for a minimum of two hours and a maximum of 24 h before limbal cell seeding.

Techniques: Fluorescence, Staining, Immunofluorescence

Journal: Stem cell research & therapy

Article Title: New characterization and safety evaluation of human limbal stem cells used in clinical application: fidelity of mitotic process and mitotic spindle morphologies.

doi: 10.1186/s13287-023-03586-z

Figure Lengend Snippet: Fig. 5 Representative flow cytometry and ddPCR analysis of LSCs and 3T3 cells prepared as the feeder layer. A and B Gating strategy of single-cell LSCs based on forward scatter (FSC) and side scatter (SSC) parameters. Unspecific staining was set to 1% of the acquired cells according to isotype antibody staining. C The majority of LSCs expressed p63α, D but few were positive for the ABCG2 stem cell-associated marker. E Residual 3T3 feeder cells in LSC expanded culture were below the acceptance limit. F 1D plot for two sets of analyses on ddPCR analysis of sample 3 (A1 – D1) and sample 4 (E1 – H1) with mouse assay. G4 is analysis of mouse positive control (DNA from pure 3T3 culture), H11 is negative control (NTC) and H12 is test for possible cross-reactivity with human DNA (human positive control). G Analysis of sample 3 (A3) and sample 4 (B3) with the human assay. H4 is analysis of human positive control (DNA from human whole blood), A11 is NTC and A12 is test for possible cross-reactivity with mouse DNA (mouse positive control)

Article Snippet: Briefly, the feeder layer was prepared from a 3T3 murine Swiss albino fibroblast cell line (ATCC® CCL-92TM, VA, USA) preferably for a minimum of two hours and a maximum of 24 h before limbal cell seeding.

Techniques: Flow Cytometry, Staining, Marker, Mouse Assay, Positive Control, Negative Control

Journal: Stem cell research & therapy

Article Title: New characterization and safety evaluation of human limbal stem cells used in clinical application: fidelity of mitotic process and mitotic spindle morphologies.

doi: 10.1186/s13287-023-03586-z

Figure Lengend Snippet: Fig. 6 Evaluation of limbal graft. A To estimate the exact time when the graft was ready for clinical use, the growth of limbal cells in the control sample on the plastic surface was monitored daily. On the first day of culture, a small colony (approximately 5 cells) of limbal stem cells (LSCs) was cocultured with a feeder layer showing undifferentiated small-sized polygonal cells with little cytoplasm (arrows). They were surrounded by 3T3 feeder layer cells, which represent larger cells, elongated irregularly shaped (branched) fibroblasts. On the second day of cultivation, a nearly circular colony of LSCs with a smooth perimeter was visible. On the third day of cultivation, circular colonies joined together (arrows), expanded and pushed away the feeder cells. On the fourth, fifth and sixth day of culture, round colonies continued to grow, forming stratifications and merged with each other until confluence with a progressively decreasing amount of feeder cells. The results of measuring the size of LSCs grown in limbal grafts (B, C) that were p63-positive cells were significantly smaller than p63-negative cells (P < 0.001) (magnification, 100 ×)

Article Snippet: Briefly, the feeder layer was prepared from a 3T3 murine Swiss albino fibroblast cell line (ATCC® CCL-92TM, VA, USA) preferably for a minimum of two hours and a maximum of 24 h before limbal cell seeding.

Techniques: Control