REVIEW pre Culture conditions for hESC

REVIEW pre Culture conditions for hESC - REPRODUCTION...

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REPRODUCTION REVIEW Focus on Stem Cells Culture conditions for human embryonic stem cells Heli Skottman 1 and Outi Hovatta 1,2 1 REGEA, Institute for Regenerative Medicine, University of Tampere and Tampere University Hospital, 33520 Tampere, Finland and 2 Division of Obstetrics and Gynecology, Department of CLINTEC, Karolinska Institutet, Karolinska University Hospital Huddinge K 57, SE 141 86 Stockholm, Sweden Correspondence should be addressed to O Hovatta; Email: [email protected] Abstract Human embryonic stem cell (hESC) lines have been derived and cultured in variable conditions. The idea behind derivation of hESC lines is to use them in human cell transplantation after differentiation, but already now these cells are widely used for research purposes. Despite similarities among the established lines, important differences have been reported between them, and it has been difficult to compare the results obtained using different lines. Recent optimization of hESC culture conditions has moved from cultures on mouse embryonic fibroblasts (MEFs) in fetal bovine serum-containing medium towards feeder-free culture methods using more defined animal substance-free cultures. The aim has been to establish robust and cost-effective systems for culturing these cells and eliminate the risk of infection transmitted byanimal pathogens and immunoreactions caused byanimal substances in cell cultures before clinical treatment. It is important to take these modifications into account when carrying out research using these cells. It is known that culture conditions influence gene expression and, hence, probably many properties of the cells. Optimization and standardization of culture methods is needed for research as well as for clinical purposes. Reproduction (2006) 132 691–698 Introduction Human embryonic stem cell (hESC) lines can be derived from the inner cell mass (ICM) of preimplantation blastocysts ( Thomson et al. 1998 ) which have been donated for research and would otherwise be discarded. Frozen good quality embryos not used in the infertility treatment, but have been used for the establishment of most of the existing lines. (hESC) Lines have been derived also form morula stage embryos ( Strelchenko et al. 2004 ), and there is a recent report stating that two hESC lines were obtained from single isolated blastomeres ( Klimanskaya et al. 2006 ), even though these blasto- meres had to be cultured in groups in order to survive. Since the establishment of the first hESC lines by Thomson et al. (1998) , rapid progress has been made and several groups have described the derivation and culture of new hESC lines in various culture conditions ( Tables 1–3 ). Human ESCs are cable of unlimited self-renewal, can be propagated in culture for extended periods and has the ability to differentiate to multiple cell types representing all primitive embryonic germ layers. Their differentiation potential has raised hope that these cells could provide new opportunities as a renewable source for cell transplantation for severe degenerative diseases.
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