Journal: RSC Advances

Article Title: Synthetic thermoresponsive scaffolds for the expansion and differentiation of human pluripotent stem cells into cardiomyocytes

doi: 10.1039/d5ra04674b

Figure Lengend Snippet: Impact of peptide incorporation on hPSC morphology and function. (A) Representative brightfield micrographs comparing WTC-11 and H9 hPSC cultures across different substrate conditions: Cultrex™, terpolymer alone (T), terpolymer with fibronectin (T + FB), and terpolymer with RGD peptide (T + RGD). Scale bars: 200 μm. (B) Cell proliferation measured as live fold expansion for both cell lines when cultured on 2 : 4 : 94 P400 terpolymer (15 wt%) modified with different bioactive molecules (FB, RGD, RGDS; 6 μg per well). (C) Maintenance of pluripotency assessed by OCT4 expression using flow cytometry under identical culture conditions. Data shown as mean ± SD with n = 5. Statistical significance: * p < 0.0001, ** p = 0.081, # p = 0.0383 compared to Cultrex™. (D) Immunofluorescence images of WTC-11 hiPSCs cultured on 2 : 4 : 94 P400 terpolymer scaffolds (15 wt%) demonstrating expression of pluripotency markers OCT-4 (green) and TRA-1-81 (red), with nuclear counterstaining using DAPI (blue). Merged images confirm maintenance of stem cell identity. Scale bars: 100 μm.

Article Snippet: Cells were cultured on Cultrex basement membrane extracts (CultrexTM) (R&D Systems, Minneapolis, USA)—a defined ECM substitute derived from EHS mouse sarcoma and used as an alternative to MatrigelTM 21 —coated 6-well plates using mTeSR-1 (StemCell Technologies, Vancouver, Canada) medium under feeder-free conditions, following a defined procedure based on Ludwig et al.

Techniques: Cell Culture, Modification, Expressing, Flow Cytometry, Immunofluorescence

Journal: RSC Advances

Article Title: Synthetic thermoresponsive scaffolds for the expansion and differentiation of human pluripotent stem cells into cardiomyocytes

doi: 10.1039/d5ra04674b

Figure Lengend Snippet: Gene expression profiles and metabolic activity of WTC-11 iPSCs and H9 hESCs cultured on the terpolymer (2 : 4 : 94 P400) with and without the addition of peptides (fibronectin (FB), RGD) at a concentration of 6 μg per well. (A) Relative expression of OCT4 assessed by qPCR for WTC-11 and H9 cells. (B) Relative expression of SOX2 assessed by qPCR for WTC-11 and H9 cells. (C) Relative expression of NANOG assessed by qPCR for H9 cells. (D) Metabolic potential (stressed OCR and ECAR) measured by Seahorse analysis for WTC-11 cells. (E) Metabolic potential (stressed OCR and ECAR) measured by Seahorse analysis for H9 cells. Cultrex™ was used as the control substrate for all comparisons. Statistical comparison between cell lines under T + RGD condition was performed to evaluate differential responses of iPSCs versus ESCs to the same scaffold modification. Statistical significance: * p < 0.03. Bars represent mean ± STDV ( n = 3 for gene expression, n = 5 for metabolic potential assays).

Article Snippet: Cells were cultured on Cultrex basement membrane extracts (CultrexTM) (R&D Systems, Minneapolis, USA)—a defined ECM substitute derived from EHS mouse sarcoma and used as an alternative to MatrigelTM 21 —coated 6-well plates using mTeSR-1 (StemCell Technologies, Vancouver, Canada) medium under feeder-free conditions, following a defined procedure based on Ludwig et al.

Techniques: Gene Expression, Activity Assay, Cell Culture, Concentration Assay, Expressing, Control, Comparison, Modification