Human mid-trimester amniotic fluid stem cells cultured under embryonic stem cell conditions with valproic acid acquire pluripotent characteristics

Dafni Moschidou, Sayandip Mukherjee, Michael P. Blundell, Gemma N. Jones, Anthony J. Atala, Adrian J. Thrasher, Nicholas M. Fisk, Paolo De Coppi, Pascale V. Guillot

Research output: Contribution to journalArticle

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Abstract

Human mid-trimester amniotic fluid stem cells (AFSC) have promising applications in regenerative medicine, being broadly multipotent with an intermediate phenotype between embryonic (ES) and mesenchymal stem cells (MSC). Despite this propluripotent phenotype, AFSC are usually cultured in adherence in a serum-based expansion medium, and how expansion in conditions sustaining pluripotency might affect their phenotype remains unknown. We recently showed that early AFSC from first trimester amniotic fluid, which endogenously express Sox2 and Klf4, can be reprogrammed to pluripotency without viral vectors using the histone deacetylase inhibitor valproic acid (VPA). Here, we show that mid-trimester AFSC cultured under MSC conditions contained a subset of cells endogenously expressing telomerase, CD24, OCT4, C-MYC, and SSEA4, but low/null levels of SOX2, NANOG, KLF4, SSEA3, TRA-1-60, and TRA-1-81, with cells unable to form embryoid bodies (EBs) or teratomas. In contrast, AFSC cultured under human ESC conditions were smaller in size, grew faster, formed colonies, upregulated OCT4 and C-MYC, and expressed KLF4 and SOX2, but not NANOG, SSEA3, TRA-1-60, and TRA-1-81. Supplementation with VPA for 5 days further upregulated OCT4, KLF4, and SOX2, and induced expression of NANOG, SSEA3, TRA-1-60, and TRA-1-81, with cells now able to form EBs and teratomas. We conclude that human mid-trimester AFSC, which may be isolated autologously during pregnancy without ethics restriction, can acquire pluripotent characteristics without the use of ectopic factors. Our data suggest that this medium-dependant approach to pluripotent mid-trimester AFSC reflects true reprogramming and not the selection of prepluripotent cells.

Original languageEnglish
Pages (from-to)444-458
Number of pages15
JournalStem Cells and Development
Volume22
Issue number3
DOIs
StatePublished - Feb 1 2013
Externally publishedYes

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Valproic Acid
Amniotic Fluid
Embryonic Stem Cells
Stem Cells
Embryoid Bodies
Teratoma
Mesenchymal Stromal Cells
Phenotype
Histone Deacetylase Inhibitors
Regenerative Medicine
Telomerase
First Pregnancy Trimester
Ethics
Pregnancy
Serum

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Hematology

Cite this

Human mid-trimester amniotic fluid stem cells cultured under embryonic stem cell conditions with valproic acid acquire pluripotent characteristics. / Moschidou, Dafni; Mukherjee, Sayandip; Blundell, Michael P.; Jones, Gemma N.; Atala, Anthony J.; Thrasher, Adrian J.; Fisk, Nicholas M.; De Coppi, Paolo; Guillot, Pascale V.

In: Stem Cells and Development, Vol. 22, No. 3, 01.02.2013, p. 444-458.

Research output: Contribution to journalArticle

Moschidou, D, Mukherjee, S, Blundell, MP, Jones, GN, Atala, AJ, Thrasher, AJ, Fisk, NM, De Coppi, P & Guillot, PV 2013, 'Human mid-trimester amniotic fluid stem cells cultured under embryonic stem cell conditions with valproic acid acquire pluripotent characteristics', Stem Cells and Development, vol. 22, no. 3, pp. 444-458. https://doi.org/10.1089/scd.2012.0267
Moschidou, Dafni ; Mukherjee, Sayandip ; Blundell, Michael P. ; Jones, Gemma N. ; Atala, Anthony J. ; Thrasher, Adrian J. ; Fisk, Nicholas M. ; De Coppi, Paolo ; Guillot, Pascale V. / Human mid-trimester amniotic fluid stem cells cultured under embryonic stem cell conditions with valproic acid acquire pluripotent characteristics. In: Stem Cells and Development. 2013 ; Vol. 22, No. 3. pp. 444-458.
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