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Dev Reprod. 2015 Sep;19(3):119-26. doi: 10.12717/DR.2015.19.3.119.

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

Author information

1
Stem Cell Research Center, Jeju National University, Jeju 63243, Korea; Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University, Jeju 63243, Korea.
2
Stem Cell Research Center, Jeju National University, Jeju 63243, Korea; Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University, Jeju 63243, Korea; Mirae Cell Bio, Seoul 05066, Korea.
3
Dept. of Preventive Medicine, College of Medicine, Jeju National University, Jeju 63243, Korea.

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.

KEYWORDS:

Feeder cell; MEF feeder cell; Mouse embryonic stem cell; Pluripotency marker

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