Effects of Feeder Cell Types on Culture of Mouse Embryonic Stem Cell In Vitro

Abstract

The suitable feeder cell layer is important for culture of embryonic stem (ES) cells. In this study, we investigated the effect of two kinds of the feeder cell, MEF cells and STO cells, layer to mouse ES (mES) cell culture for maintenance of stemness. We compare the colony formations, alkaline phosphatase (AP) activities, expression of pluripotency marker genes and proteins of D3 cell colonies cultured on MEF feeder cell layer (D3/MEF) or STO cell layers (D3/STO) compared to feeder free condition (D3/–) as a control group. Although there were no differences to colony formations and AP activities, interestingly, the transcripts level of pluripotency marker genes, Pou5f1 and Nanog were highly expressed in D3/MEF (79 and 93) than D3/STO (61and 77) or D3/– (65 and 81). Also, pluripotency marker proteins, NANOG and SOX-2, were more synthesized in D3/MEF (72.8±7.69 and 81.2±3.56) than D3/STO (32.0±4.30 and 56.0±4.90) or D3/– (55.0±4.64 and 62.0±6.20). These results suggest that MEF feeder cell layer is more suitable to mES cell culture.

Visualization of morphological categories of colonies for monitoring of effect on induced pluripotent stem cell culture status

“…In this analysis, image-based colony morphology analysis was applied to evaluate the influence of stressful passage on feeder cells. The condition of feeder cells is considered to be an influential factor that can change the quality of iPSCs even under the use of same protocol [18] , [19] , [20] . As a model, the same conditioned iPSCs on two types of feeder cell conditions were designed to mimic the differences of feeder cell preparation skills: (i) Feeder-P3, which only experienced 3 passages after thawing of purchased cryo-vial, and (ii) Feeder-P11, which intentionally continued extra 8-times of passages of Feeder-P3.…”

“…2, the influence of feeder-cell preparation skill was evaluated. Feeder cells are known to influence the quality of PSCs [18] , [19] , therefore early passage of cells are commonly recommended. However, the definition of “early” had been ambiguous.…”

Human Pluripotent Stem Cell Culture: Current Status, Challenges, and Advancement

“…They also reported the need for selenium in media to support sustained expansion and transferrin to improve survival and cloning efficiency [41]. Selenium is also important in culture media as it has been shown to protect cells against oxidative damage by optimizing the activity of glutathione peroxidase and thioredoxin reductase, in addition to stimulating cell growth and proliferation [43].…”

Effects of in vitro Culture Period of Reconstructed Embryos and Genetic Background of Feeder Cells on Establishment of Embryonic Stem Cells Derived from Somatic Cell Nuclear Transfer Blastocysts in Pigs

“…Feeder cells are routinely used as an attachment matrix for ESC establishment and maintenance (Llames et al, 2015;Siriboon et al, 2015) because they secrete unknown factors that support proliferation and inhibit differentiation (Yang et al, 2016;Li et al, 2017). Because these unknown factors secreted from feeder cells differ according to the genetic background of feeder cells (Talbot et al, 2012), preferred genetic background of feeder cells is dependent on ESC lines (Brevini et al, 2012;Park et al, 2015;Yang et al, 2016). Therefore, optimization of genetic background of feeder cell is also essential for suc-…”