Park, Yun-Gwi
(Stem Cell Research Center, Jeju National University)
,
Lee, Seung-Eun
(Stem Cell Research Center, Jeju National University)
,
Kim, Eun-Young
(Stem Cell Research Center, Jeju National University)
,
Hyun, Hyuk
(Stem Cell Research Center, Jeju National University)
,
Shin, Min-Young
(Stem Cell Research Center, Jeju National University)
,
Son, Yeo-Jin
(Stem Cell Research Center, Jeju National University)
,
Kim, Su-Young
(Dept. of Preventive Medicine, College of Medicine, Jeju National University)
,
Park, Se-Pill
(Stem Cell Research Center, Jeju National University)
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 phosp...
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 ($72.8{\pm}7.69$ and $81.2{\pm}3.56$) than D3/STO ($32.0{\pm}4.30$ and $56.0{\pm}4.90$) or D3/- ($55.0{\pm}4.64$ and $62.0{\pm}6.20$). These results suggest that MEF feeder cell layer is more suitable to mES cell culture.
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 ($72.8{\pm}7.69$ and $81.2{\pm}3.56$) than D3/STO ($32.0{\pm}4.30$ and $56.0{\pm}4.90$) or D3/- ($55.0{\pm}4.64$ and $62.0{\pm}6.20$). These results suggest that MEF feeder cell layer is more suitable to mES cell culture.
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제안 방법
To check the phenotypic pluripotency, we performed AP assay with D3 cell colonies on MEF or STO feeder cell layer. AP activities presented violet color under a visible ray were highly detected in D3/MEF, D3/STO and D3/– and there was no differences among groups.
대상 데이터
In this study, we compared the effects of two kind feeder cells, MEF cell and STO cell, on maintenance of stemness in mouse ES (mES) cell. The ES-D3 (D3) cell line, one of the well-known mES cell line, is used. We checked colony formation, AP activity, identification of pluripotency markers of D3 cell colonies.
데이터처리
A paired Student’s t-test was used to compare relative gene expression.
이론/모형
The general linear model (GLM) procedure within the Statistical Analysis System (SAS User’s Guide, 1985, Statistical Analysis System Inc., Cary, NC. USA) was used to analyze data from all experiments.
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