Journal: Cancers

Article Title: The Potential of Human Pulmonary Mesenchymal Stem Cells as Vectors for Radiosensitizing Metallic Nanoparticles: An In Vitro Study †

doi: 10.3390/cancers16183239

Figure Lengend Snippet: Dose- and time-dependent effects of Fe 3 O 4 @Au nanoparticles on the viability of HPMSCs. Cells were incubated with increasing concentrations of Fe 3 O 4 @Au NPs (10–500 µg/mL). ( A ) The mitochondrial metabolic activity of HPMSCs was quantified via an MTT assay after 24 h, ( B ) 48 h, and ( C ) 72 h of incubation with Fe 3 O 4 @Au NPs, and ( D ) cell counting was performed via the trypan blue exclusion method on cells after 24 h, ( E ) 48 h, or ( F ) 72 h of incubation with Fe 3 O 4 @Au NPs. Data are expressed as the mean ± SEM of three independent experiments. *: p < 0.05, **: p < 0.01, ***: p < 0.001, and ****: p < 0.0001 as compared to control conditions.

Article Snippet: Primary human pulmonary mesenchymal stem cells (HPMSCs) isolated from human lung tissue were purchased from the ScienCell research laboratory (Carlsbad, CA, USA) and were cultured in Modified Eagle’s Medium (MEM; PAN Biotech, Aidenbach, Germany) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech), 1 mM of sodium pyruvate (PAN Biotech), 0.5 μg/mL of amphotericin B (PAN Biotech), 2 mM of L-glutamine (Biochrom AG, Berlin, Germany), and 0.1 mg/mL penicillin–streptomycin (PAN Biotech).

Techniques: Incubation, Activity Assay, MTT Assay, Cell Counting, Control

Journal: Cancers

Article Title: The Potential of Human Pulmonary Mesenchymal Stem Cells as Vectors for Radiosensitizing Metallic Nanoparticles: An In Vitro Study †

doi: 10.3390/cancers16183239

Figure Lengend Snippet: Evaluation of the cellular uptake of Fe 3 O 4 @Au nanoparticles by HPMSCs. Cellular internalization was assessed using Prussian blue staining. The internalization of Fe 3 O 4 @Au NPs was determined on ( A ) control cells and ( B ) cells exposed to 100 µg/mL Fe 3 O 4 @Au NPs for 24 h, as well as ( C ) control cells and ( D ) cells exposed to 100 µg/mL Fe 3 O 4 @Au for 72 h. Blue staining is consistent with the intracellular uptake of Fe 3 O 4 @Au NPs by HPMSCs.

Article Snippet: Primary human pulmonary mesenchymal stem cells (HPMSCs) isolated from human lung tissue were purchased from the ScienCell research laboratory (Carlsbad, CA, USA) and were cultured in Modified Eagle’s Medium (MEM; PAN Biotech, Aidenbach, Germany) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech), 1 mM of sodium pyruvate (PAN Biotech), 0.5 μg/mL of amphotericin B (PAN Biotech), 2 mM of L-glutamine (Biochrom AG, Berlin, Germany), and 0.1 mg/mL penicillin–streptomycin (PAN Biotech).

Techniques: Staining, Control

Journal: Cancers

Article Title: The Potential of Human Pulmonary Mesenchymal Stem Cells as Vectors for Radiosensitizing Metallic Nanoparticles: An In Vitro Study †

doi: 10.3390/cancers16183239

Figure Lengend Snippet: Effect of Fe 3 O 4 @Au nanoparticles on the necrosis and apoptosis markers of HPMSCs. ( A ) The cytotoxic effect of 100 µg/mL Fe 3 O 4 @Au NPs on cells was assessed using an LDH assay after 24 h and ( B ) 72 h of exposure to nanoparticles. The mean expression of genes encoding ( C ) caspase 3, ( D ) caspase 9, and ( E ) Bcl2 was obtained via RT-qPCR on HPMSCs incubated or not with Fe 3 O 4 @Au NPs for 24 h and 72 h. Data are indicated as the mean ± SEM of three independent experiments. *: p < 0.05, **: p < 0.01, and ****: p < 0.0001 as compared to the corresponding control conditions.

Article Snippet: Primary human pulmonary mesenchymal stem cells (HPMSCs) isolated from human lung tissue were purchased from the ScienCell research laboratory (Carlsbad, CA, USA) and were cultured in Modified Eagle’s Medium (MEM; PAN Biotech, Aidenbach, Germany) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech), 1 mM of sodium pyruvate (PAN Biotech), 0.5 μg/mL of amphotericin B (PAN Biotech), 2 mM of L-glutamine (Biochrom AG, Berlin, Germany), and 0.1 mg/mL penicillin–streptomycin (PAN Biotech).

Techniques: Lactate Dehydrogenase Assay, Expressing, Quantitative RT-PCR, Incubation, Control

Journal: Cancers

Article Title: The Potential of Human Pulmonary Mesenchymal Stem Cells as Vectors for Radiosensitizing Metallic Nanoparticles: An In Vitro Study †

doi: 10.3390/cancers16183239

Figure Lengend Snippet: Effect of Fe 3 O 4 @Au nanoparticles on the redox status of HPMSCs. The mean normalized expression of genes encoding redox markers was determined by RT-qPCR. The expressions of genes coding for ( A ) Nox4, ( B ) HMOX-1, and ( C ) Nrf2 in HPMSCs were measured after 24 h and 72 h of exposure to 100 µg/mL Fe 3 O 4 @Au NPs. ( D ) ROS production was determined in HPMSCs exposed to 100 µg/mL Fe 3 O 4 @Au NPs for 2 h by using the DCFH-DA probe. Data are expressed as the mean ± SEM of three independent experiments. **: p < 0.01 and ****: p < 0.0001 as compared to the corresponding control conditions.

Article Snippet: Primary human pulmonary mesenchymal stem cells (HPMSCs) isolated from human lung tissue were purchased from the ScienCell research laboratory (Carlsbad, CA, USA) and were cultured in Modified Eagle’s Medium (MEM; PAN Biotech, Aidenbach, Germany) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech), 1 mM of sodium pyruvate (PAN Biotech), 0.5 μg/mL of amphotericin B (PAN Biotech), 2 mM of L-glutamine (Biochrom AG, Berlin, Germany), and 0.1 mg/mL penicillin–streptomycin (PAN Biotech).

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Cancers

Article Title: The Potential of Human Pulmonary Mesenchymal Stem Cells as Vectors for Radiosensitizing Metallic Nanoparticles: An In Vitro Study †

doi: 10.3390/cancers16183239

Figure Lengend Snippet: Effect of Fe 3 O 4 @Au nanoparticles on proinflammatory cytokines and chemokines secreted by HPMSCs. The proinflammatory response of HPMSCs was evaluated through ELISA. The levels of ( A ) TNF-α, ( B ) IL-1β, ( C ) IL-6, ( D ) CXCL8, and ( E ) CCL5 secreted by HPMSCs were quantified after 24 or 72 h of exposure to 100 µg/mL of Fe 3 O 4 @Au NPs. Data are presented as the mean ± SEM of three independent experiments. *: p < 0.05, **: p < 0.01, ***: p < 0.001, and ****: p < 0.0001 as compared to the corresponding control condition.

Article Snippet: Primary human pulmonary mesenchymal stem cells (HPMSCs) isolated from human lung tissue were purchased from the ScienCell research laboratory (Carlsbad, CA, USA) and were cultured in Modified Eagle’s Medium (MEM; PAN Biotech, Aidenbach, Germany) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech), 1 mM of sodium pyruvate (PAN Biotech), 0.5 μg/mL of amphotericin B (PAN Biotech), 2 mM of L-glutamine (Biochrom AG, Berlin, Germany), and 0.1 mg/mL penicillin–streptomycin (PAN Biotech).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Cancers

Article Title: The Potential of Human Pulmonary Mesenchymal Stem Cells as Vectors for Radiosensitizing Metallic Nanoparticles: An In Vitro Study †

doi: 10.3390/cancers16183239

Figure Lengend Snippet: Effect of Fe 3 O 4 @Au nanoparticles on the expression and production of protumorigenic factors by the HPMSCs. The gene expression and secreted levels of protumorigenic markers were measured on cells exposed to 100 µg/mL Fe 3 O 4 @Au nanoparticles for 24 and 72 h. First, the mean expression of genes encoding ( A ) PI3K, ( B ) AMPK, ( C ) PDGF, and ( D ) VEGF were determined via the use of RT-qPCR. Then, ( E ) the level of VEGF secreted by HPMSCs was quantified through an ELISA. Data are expressed as the mean ± SEM of three independent experiments. *: p < 0.05 and ****: p < 0.0001 as compared to the corresponding control conditions.

Article Snippet: Primary human pulmonary mesenchymal stem cells (HPMSCs) isolated from human lung tissue were purchased from the ScienCell research laboratory (Carlsbad, CA, USA) and were cultured in Modified Eagle’s Medium (MEM; PAN Biotech, Aidenbach, Germany) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech), 1 mM of sodium pyruvate (PAN Biotech), 0.5 μg/mL of amphotericin B (PAN Biotech), 2 mM of L-glutamine (Biochrom AG, Berlin, Germany), and 0.1 mg/mL penicillin–streptomycin (PAN Biotech).

Techniques: Expressing, Gene Expression, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Control

Journal: Cell Cycle

Article Title: Context-dependent regulation of Musashi-mediated mRNA translation and cell cycle regulation

doi: 10.4161/cc.10.1.14388

Figure Lengend Snippet: Musashi activates mRNA translation in differentiating neural stem/progenitor cells. Primary neural stem/progenitor cells (NSPCs) were cultured from embryonic rat hippocampal/cortical tissue (Genlantis, San Diego) and proliferation of the NSPCs established through assay of neurosphere formation.35 Differentiation was induced by plating enzymatically and mechanically dispersed single cells on poly-ornithine/fibronectin-coated dishes in the absence of bFGF and EGF and in the presence of retinoic acid.36 Dispersed single NSPCs were co-transfected with plasmids encoding Renilla luciferase and either a Firefly luciferase fused to either a control 3′ UTR (Con) or a 3′ UTR containing a MBE (MBE). Cells were then cultured in either proliferative (Prolif) or differentiation-inducing (Diff) media. 24 hours after the transfection, the cells were lysed and samples prepared for both luciferase activity and reporter mRNA expression as described in the legend to Figure 3. (A) Luciferase values are shown relative to the Firefly luciferase fused to the control 3′ UTR, arbitrarily set to 100% for both proliferation and differentiation conditions (Prolif/Diff). Error bars represent SEM from three independent experiments. Repression of the Fluc-MBE reporter in proliferating NSPCs was significant (p < 0.005, Student's t-test) as was the activation of the Fluc-MBE reporter in differentiating NSPCs (p < 0.05, Student's t-test). (B) The levels of Firefly reporter mRNA were determined using semi-quantitative PCR and no significant differences in stability of the Fluc-Con or Fluc-MBE reporter mRNAs were detected in NSPCs. The Fluc-MBEmt reporter mRNA gave results indistinguishable from the Fluc-Con reporter (data not shown). m, indicates DNA marker lane.

Article Snippet: Primary neural stem/progenitor cells (NSPCs) were cultured from embryonic rat hippocampal/cortical tissue (Genlantis, San Diego) and proliferation of the NSPCs established through assay of neurosphere formation.

Techniques: Cell Culture, Transfection, Luciferase, Activity Assay, Expressing, Activation Assay, Real-time Polymerase Chain Reaction, Marker

Journal: Advanced Science

Article Title: The Bioengineering of Microspheric Skin Organoids and Their Application in Drug Screening

doi: 10.1002/advs.202416863

Figure Lengend Snippet: Fabrication of microspheric skin organoids. A). A schematic diagram (Created in BioRender) illustrating the formation of microspheric skin organoids. B). Photographs of human dermal fibroblast (HDF)‐spheres at day 0 and day 1 under a light microscopy. C). Representative images of day 5 microspheric skin organoids (left), and the cross section of the organoid after immunofluorescent staining for E‐cadherin (E‐cad) and vimentin. D). Representative images of cross‐sections of a day 7 organoid after immunofluorescence staining for F‐actin and E‐cad. E). Representative microscopic images of a day 7 microspheric skin organoid (whole mount) showing a core‐shell structure, with the core composed of mCherry‐expressing HDFs (red) and the shell formed by GFP‐expressing HaCaT cells (green). Scale bar: 100 µm.

Article Snippet: HaCaT cells, human foreskin primary epidermal stem cells (Epi‐SCs), and human dermal fibroblasts (HDFs) were purchased from iCell Bioscience Co., Ltd (Shanghai).

Techniques: Light Microscopy, Staining, Immunofluorescence, Expressing

Journal: Advanced Science

Article Title: The Bioengineering of Microspheric Skin Organoids and Their Application in Drug Screening

doi: 10.1002/advs.202416863

Figure Lengend Snippet: Terminal differentiation of the epidermis in microspheric skin organoids. A). A schematic diagram (Created in BioRender) illustrating the formation of cornified microspheric skin organoids. Organoids consisting of primary human keratinocytes and HDFs were first cultured in growth medium to allow keratincoytes to proliferate, and then in induction medium to induce epidermal maturation. B). Representative images of microspheric skin organoids prepared by different keratinocytes. Cytokeratins were stained using a pan‐cytokeratin (Pan‐CK) antibody, E‐cadherin (E‐cad) was stained in red, and nuclei were stained blue. C). Lucifer Yellow permeability assay of the human dermal fibroblast core (HDF, left), skin organoids with epidermis formed by HaCaT cells (middle), or by primary human keratinocytes (right), where the dye was in green color and the nuclei were in blue. D). Sections of day 7 organoids comprising primary human keratinocytes were stained for keratin 5, keratin 10, and filaggrin to illustrate the differentiation of the epidermal cells. Scale bars: 100 µm.

Article Snippet: HaCaT cells, human foreskin primary epidermal stem cells (Epi‐SCs), and human dermal fibroblasts (HDFs) were purchased from iCell Bioscience Co., Ltd (Shanghai).

Techniques: Cell Culture, Staining, Permeability