Available human feeder cells for the maintenance of human embryonic stem cells

Abstract

Mouse embryonic fibroblasts (MEFs) have been previously used as feeder cells to support the growth of human embryonic stem cells (hESCs). In this study, human adult uterine endometrial cells (hUECs), human adult breast parenchymal cells (hBPCs) and embryonic fibroblasts (hEFs) were tested as feeder cells for supporting the growth of hESCs to prevent the possibility of contamination from animal feeder cells. Cultured hUECs, hBPCs and hEFs were mitotically inactivated and then plated. hESCs (Miz-hES1, NIH registered) initially established on mouse feeder layers were transferred onto each human feeder layer and split every 5 days. The morphology, expression of specific markers and differentiation capacity of hESCs adapted on each human feeder layer were examined. On hUEC, hBPC and hEF feeder layers, hESCs proliferated for more than 90, 50 and 80 passages respectively. Human feeder-based hESCs were positive for stage-specific embryonic antigen (SSEA)-3 and -4, and Apase; they also showed similar differentiation capacity to MEF-based hESCs, as assessed by the formation of teratomas and expression of tissue-specific markers. However, hESCs cultured on hUEC and hEF feeders were slightly thinner and flatter than MEF-or hBPC-based hESCs. Our results suggest that, like MEF feeder layers, human feeder layers can support the proliferation of hESCs without differentiation. Human feeder cells have the advantage of supporting more passages than when MEFs are used as feeder cells, because hESCs can be uniformly maintained in the undifferentiated stage until they pass through senescence. hESCs established and/or maintained under stable xeno-free culture conditions will be helpful to cell-based therapy.

Designer blood: creating hematopoietic lineages from embryonic stem cells

“…[12][13][14] In this regard, further studies to optimize the propagation of human ES cells and eliminate their requirements for animal feeder cells are important for future clinical applications. For example, human ES cells can be maintained on human stroma, 15,16 and numerous signaling pathways and transcription factors that participate in ES-cell self-renewal have been exploited for developing feeder-free culture systems. [17][18][19][20] This includes the use of b-FGF 17,21 and Wnt signaling agonists.…”

Analysis of the Factors that Limit the Ability of Feeder Cells to Maintain the Undifferentiated State of Human Embryonic Stem Cells

“…Human ESCs (hESCs) were derived and cultured initially on feeder cell layers (FLs), composed of mitotically inactivated (irradiated) mouse embryonic fi broblasts (MEFs) [1]. Although the use of MEF as FL has become the conventional method of hESC culture, human fetal and adult fi broblasts have also been used successfully for hESC culture [2][3][4][5], and autogenic FL systems have also been developed [6].…”

Stem Cells and Scaffolds for Vascularizing Engineered Tissue Constructs

“…Successful animal-free culture systems, however, have been developed that remove the need for nonhuman feeder layers and serum environments. In these studies, hESCs, cultured on a number of different human feeder layers, including human embryonic fibroblasts, adult fallopian tube epithelial cells, and human foreskins, have shown the ability to maintain hESC-pluripotency, immortality, and proliferative capabilities [122][123][124] . An alternative is to forego feeder layers and use a serum-free environment, attained by using serum replacement in combination with a variety of cytokines.…”