Effect of Passage, Sorting, and Media on Differentiation Capacity and Marker Expression in Amniotic Fluid Stem Cells

Jennifer P. Connell, Emily Augustini, Shannon Cheng, Omar M. Benavides, Rodrigo Ruano, Jeffrey G. Jacot

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Amniotic fluid-derived stem cells (AFSC) represent a promising source for tissue engineering applications; however, wide variation exists in the methods and medium used in AFSC culture. To address the maintenance of stem cell markers and differentiation capacity of AFSC, both c-kit sorted and unfractionated adherent AFSC from human patient samples were cultured in a variety of media and evaluated over six passages. Medium supplements included Chang medium, basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and both bFGF and EGF. Protein and gene expression of pluripotency markers, including c-kit, SSEA4, Sox2, Oct4, and cMyc were measured in undifferentiated AFSC. Differentiation capacities of AFSC towards endothelial, osteogenic, and neurogenic lineages were analyzed at passages 5 and 6. Expression of c-kit was higher in c-kit sorted cells compared to unfractionated cells; however, SSEA4, Sox2, cMyc, Tra-1-60, and Tra-1-81 were higher in the unfractionated population. Pluripotency marker expression was significantly lower at passage 6 than earlier passages. Correspondingly, cells differentiated more efficiently at passage 5 than passage 6. Media had distinct effects on differentiation towards each lineage. These results indicate that AFSC should be utilized prior to passage 6 and that optimal isolation and culture conditions depend on final differentiation intent.

Original languageEnglish (US)
Pages (from-to)139-150
Number of pages12
JournalCellular and Molecular Bioengineering
Volume9
Issue number1
DOIs
StatePublished - Mar 1 2016
Externally publishedYes

Fingerprint

Stem Cells
Differentiation Antigens
Amniotic Fluid
Stem cells
Sorting
Fluid
Fluids
Growth Factors
Fibroblasts
Fibroblast Growth Factor 2
Epidermal Growth Factor
Cell
Tissue Engineering
Cell Culture
Engineering Application
Tissue engineering
Cell culture
Gene expression
Isolation
Gene Expression

Keywords

  • Amnion
  • C-kit
  • Multipotential differentiation
  • Perinatal stem cells

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Modeling and Simulation

Cite this

Effect of Passage, Sorting, and Media on Differentiation Capacity and Marker Expression in Amniotic Fluid Stem Cells. / Connell, Jennifer P.; Augustini, Emily; Cheng, Shannon; Benavides, Omar M.; Ruano, Rodrigo; Jacot, Jeffrey G.

In: Cellular and Molecular Bioengineering, Vol. 9, No. 1, 01.03.2016, p. 139-150.

Research output: Contribution to journalArticle

Connell, Jennifer P. ; Augustini, Emily ; Cheng, Shannon ; Benavides, Omar M. ; Ruano, Rodrigo ; Jacot, Jeffrey G. / Effect of Passage, Sorting, and Media on Differentiation Capacity and Marker Expression in Amniotic Fluid Stem Cells. In: Cellular and Molecular Bioengineering. 2016 ; Vol. 9, No. 1. pp. 139-150.
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