Culture conditions for human embryonic stem cells

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 by animal pathogens and immunoreactions caused by animal 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.

A Survey of Parameters Involved in the Establishment of New Lines of Human Embryonic Stem Cells

“…In agreement to this, FBS has been shown to promote differentiation of the ESC at a higher rate [35]. Furthermore, although containing animal-derived components, KSR is a defined supplement, avoiding the lot-to-lot composition variation found in FBS [36]. Nevertheless, even in the last 2 years, a significant number of hESC lines (21 lines out of 91 informative lines-23.1%) were still established in a combination of KSR/FBS, which apparently improves the initial outgrowth of the ICM, and then are subsequently transitioned into KSR alone for long term culture [17] (Fig.…”

Engineering integrin signaling for promoting embryonic stem cell self-renewal in a precisely defined niche

“…Several attempts have been described to replace MEF feeder layers with ECM proteins and matrices such as Matrigel, ECM derived from MEFs, etc. [1][2][3]. These biologically derived materials are also very complex and are not subject to control at the molecular level.…”

Use of Cytogenetic Analysis Methods for Assessing the Quality of Cell Lines in Biomedical Cell Products

“…Последнее свойство присуще также и опу-холевым клеткам. Существует много работ, подтверждающих наличие значительных генетических изменений ЭСК при дли-тельном культивировании [36][37][38]. В медицинской практике были зарегистрированы случаи развития опухолей у пациентов при трансплантации ЭСК [39].…”