Proliferation and Pluripotency of Human Embryonic Stem Cells Maintained on Type I Collagen

Abstract

Human embryonic stem cells (hESC) require a balance of growth factors and signaling molecules to proliferate and retain pluripotency. Conditioned medium (CM) from a human embryonic germ-cell-derived cell culture, SDEC, was observed to support the growth of hESC on type I collagen (COL I) and on Matrigel (MAT) biomatricies. After 1 month, the population doubling of hESC grown in SDEC CM on COL I was equivalent to that of hESC grown in mouse embryonic fibroblast (MEF) CM on MAT. hESC grown in SDEC CM on COL I expressed OCT4, NANOG, SSEA-4, alkaline phosphatase (AP), and TRA-1-60; retained a normal karyotype; and were capable of forming teratomas. DNA microarray analysis was used to compare the transcriptional profiles of SDEC and the less supportive WI38 and Detroit 551 human cell lines. The mRNA level of secreted frizzledrelated protein (sFRP-1), a known antagonist of the WNT=b-catenin signaling pathway, was significantly reduced in SDEC as compared with the other 2 cell lines, whereas the mRNA levels of prostaglandin-endoperoxide synthase 2 (PTGS2 or COX-2) and prostaglandin I 2 synthase (PGIS), two prostaglandin biosynthesis genes, were significantly increased in SDEC. The level of sFRP-1 protein was significantly reduced, and levels of 2 prostaglandins that are downstream products of PTGS2 and PGIS, prostaglandin E 2 and 6-keto-prostaglandin F 1a , were significantly elevated in SDEC CM compared with WI38, Detroit 551, and MEF CM. Further, addition of purified sFRP-1 to SDEC CM reduced the proliferation of hESC grown on COL I as well as MAT in a dosedependent manner.

Effects of High-Mobility Group A Protein Application on Canine Adipose-Derived Mesenchymal Stem CellsIn Vitro

“…The key factor in terms of multipotency and self-renewal characteristics of stem cells is the control by the expression of several unique multipotency-associated marker genes as for example, KLF4, cMYC, SOX2, HMGA2, and OCT4 [13–16, 40]. …”

Collagen I regulates the self-renewal of mouse embryonic stem cells through α2β1 integrin- and DDR1-dependent Bmi-1

“…These results provide evidence of a strong connection between ECM and maintenance of stem cell characteristics. Another study reported that ECSs maintained in the presence of collagen I have superior proliferation rates and pluripotency potential (Jones et al, 2010). Moreover, the present results show that collagen I (10 µg/ml) maintained the mESCs in an undifferentiated state in the presence of LIF.…”

ECM-Incorporated Hydrogels Cross-Linked via Native Chemical Ligation To Engineer Stem Cell Microenvironments

“…We sought to utilize two different families of ECM proteins (fibrillar Col I and reticular Laminin) to establish proof of concept. In addition, we selected these two molecules as they have both been included in protocols designated either to enhance stem cell growth 61,62 or to encourage stem cell differentiation to cardiomyocytes, 63–66 which is of particular interest for our laboratory. Previously, we reported that Col I and Laminin are two of the primary constituents of the developing myocardium.…”