Proliferation and pluripotency of human embryonic stem cells maintained on type i collagen

Meredith B. Jones, Chia H. Chu, James C. Pendleton, Michael J. Betenbaugh, Joseph Shiloach, Bolormaa Baljinnyam, Jeffrey S. Rubin, Michael J. Shamblott

Research output: Contribution to journalArticle

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 frizzled-related protein (sFRP-1), a known antagonist of the WNT/β-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 I2 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 E2 and 6-keto-prostaglandin F, 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 dose-dependent manner.

Original languageEnglish (US)
Pages (from-to)1923-1935
Number of pages13
JournalStem Cells and Development
Volume19
Issue number12
DOIs
StatePublished - Dec 1 2010

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Conditioned Culture Medium
Collagen
Cyclooxygenase 2
Epoprostenol
Prostaglandin-Endoperoxide Synthases
Prostaglandins
Fibroblasts
Cell Line
Catenins
Messenger RNA
Teratoma
Microarray Analysis
Collagen Type I
Human Embryonic Stem Cells
Oligonucleotide Array Sequence Analysis
Karyotype
Dinoprostone
Alkaline Phosphatase
Intercellular Signaling Peptides and Proteins
Cell Culture Techniques

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Hematology

Cite this

Jones, M. B., Chu, C. H., Pendleton, J. C., Betenbaugh, M. J., Shiloach, J., Baljinnyam, B., ... Shamblott, M. J. (2010). Proliferation and pluripotency of human embryonic stem cells maintained on type i collagen. Stem Cells and Development, 19(12), 1923-1935. https://doi.org/10.1089/scd.2009.0326

Proliferation and pluripotency of human embryonic stem cells maintained on type i collagen. / Jones, Meredith B.; Chu, Chia H.; Pendleton, James C.; Betenbaugh, Michael J.; Shiloach, Joseph; Baljinnyam, Bolormaa; Rubin, Jeffrey S.; Shamblott, Michael J.

In: Stem Cells and Development, Vol. 19, No. 12, 01.12.2010, p. 1923-1935.

Research output: Contribution to journalArticle

Jones, MB, Chu, CH, Pendleton, JC, Betenbaugh, MJ, Shiloach, J, Baljinnyam, B, Rubin, JS & Shamblott, MJ 2010, 'Proliferation and pluripotency of human embryonic stem cells maintained on type i collagen', Stem Cells and Development, vol. 19, no. 12, pp. 1923-1935. https://doi.org/10.1089/scd.2009.0326
Jones MB, Chu CH, Pendleton JC, Betenbaugh MJ, Shiloach J, Baljinnyam B et al. Proliferation and pluripotency of human embryonic stem cells maintained on type i collagen. Stem Cells and Development. 2010 Dec 1;19(12):1923-1935. https://doi.org/10.1089/scd.2009.0326
Jones, Meredith B. ; Chu, Chia H. ; Pendleton, James C. ; Betenbaugh, Michael J. ; Shiloach, Joseph ; Baljinnyam, Bolormaa ; Rubin, Jeffrey S. ; Shamblott, Michael J. / Proliferation and pluripotency of human embryonic stem cells maintained on type i collagen. In: Stem Cells and Development. 2010 ; Vol. 19, No. 12. pp. 1923-1935.
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