Human serum as an alternative for amniotic fluid stem cells culture media for clinical use

Abstract

Objectiveto examine growth, phenotypic characteristics, gene expression and differentiation of amniotic fluid stem cells (AFSC) using human serum in culture media.IntroductionAFSC are promising candidates for cell‐based therapies. They are obtained by prenatal screening, without raising the ethical concerns associated with human embryonic research. One major obstacle for their clinical use is the biosafety of fetal calf serum (FCS).MethodsAmniotic fluid cells obtained from 2nd trimester amniocentesis were grown in α‐MEM+human serum20%. Attached cells were cultured and counted under microscope, investigated about the expression of Oct‐4, SOX2 and Nanog, by RT‐PCR and about phenotype by FACS‐analysis. Induced osteogenic and chondrogenic differentiation were confirmed with Alizarine Red and H&E stains respectively.ResultsAFSC showed high proliferation ratios; expressed Oct‐4, SOX2 and Nanog; stained positive for CD29, CD44, CD90, CD105 and negative for CD14, CD31, CD34, CD45, CD106 and CD133; showed osteogenic and chondrogenic potential.ConclusionsGrowth of AFSC in a FCS‐free medium is feasible. Can be isolated easily, proliferate quickly, show a mesenchymal phenotype and have osteogenic and chondrogenic potential. Moreover, Oct‐4, SOX2 and Nanog, expressed by AFSC, are associated with the pluripotency and for the maintenance of stem cells.