Effect of extracellular matrix proteins on the differentiation of human pluripotent stem cells into mesenchymal stem cells

Abstract

The transplantation of human mesenchymal stem cells (hMSCs), such as bone marrow stem cells (BMSCs) and adipose-derived stem cells (ADSCs), has shown beneficial effects in protecting transplanted tissues and cells...

Designed peptide-grafted hydrogels for human pluripotent stem cell culture and differentiation

“…The embryoid body (EB) formation of hPSCs was performed following a conventional method. 20,26,40,41 After EB formation from hPSCs cultivated on PVAI hydrogels conjugated with designed peptides for 10 passages, the cells were analyzed by immunohistochemical staining utilizing antibodies against markers of three embryonic germline layers (glial fibrillary acidic protein (GFAP, ectoderm), smooth muscle actin (SMA, mesoderm) and α-fetoprotein (AFP, endoderm)) using a conventional method, and the differentiated cells were analysed. 20,26,40,41…”

“…Teratoma formation assays were performed following a conventional method 20,26,40,41,47,48 after hPSCs were cultured on PVAI hydrogels conjugated with designed peptides for 10 passages. Briefly, after 6–8 weeks of subcutaneously transplanting hPSCs into nonobese diabetic/severe combined immunodeficiency (NOD.CB17-Prkdcscid/JNarl NOD/SCID) mice, teratomas were commonly generated, isolated and fixed in a formaldehyde solution.…”

“…Following a conventional protocol, the sliced teratomas were subjected to hematoxylin and eosin (H&E) staining. 20,26,40,41…”

“…On day 1, the media were exchanged into Essential 6 (E6) media containing BMP4 (10 ng mL −1 ) and A83-01 (1 μM). 26 On day 3, the media were exchanged into MSC media (α-MEM containing 1% antibiotic-antimycotic and 20% fetal bovine serum (FBS)), and the cells were cultivated for over 60 days, after which the media were exchanged into fresh hMSC media once every two days. The cells were passaged utilizing a conventional method with trypsin-EDTA solution until the cell confluence reached a value higher than 85% (typically five to seven days).…”

“…The cells were passaged utilizing a conventional method with trypsin-EDTA solution until the cell confluence reached a value higher than 85% (typically five to seven days). 26…”