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Comparative Analysis for In Vitro Differentiation Potential of Induced Pluripotent Stem Cells, Embryonic Stem Cells, and Spermatogonial Stem Cells into Germ-lineage Cells

이현정|조중현|김형준|도정태|이동율|고정재|고영은
한국발생생물학회 (The Korea Society Of Developmental Biology)|2011.03|발생과 생식 (DEVELOPMENT & REPRODUCTION)|제15권 제1호|pp. 41-52

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키워드

Induced pluripotent stem cellsMouse embryonic stem cellsMultipotent spermatogonial stem cellsGerm cell lineageDifferentiation potential.
주제분류생물학

초록

In the present study, embryoid bodies (EBs) obtained from induced pluripotent stem cells (iPSCs) were induced to differentiate into germ lineage cells by treatment with bone morphogenetic protein 4 (BMP4) and retinoic acid (RA). The results were compared to the results for embryonic stem cells (ESCs) and multipotent spermatogonial stem cells (mSSCs) and quantified using immunocytochemical analysis of germ cell-specific markers (integrin-α6, GFR-α1, CD90/ Thy1), fluorescence activating cell sorting (FACS), and real time-RT-PCR. We show that the highest levels of germ cell marker-expressing cells were obtained from groups treated with 10 ng/㎖ BMP4 or 0.01 μM RA. In the BMP4-treated group, GFR-α1 and CD90/Thy-1 were highly expressed in the EBs of iPSCs and ESCs compared to EBs of mSSCs. The expression of Nanog was much lower in iPSCs compared to ESCs and mSSCs. In the RA treated group, the level of GFR-α1 and CD90/Thy-1 expression in the EBs of mSSCs was much higher than the levels found in the EBs of iPSCs and similar to the levels found in the EBs of ESCs. FACS analysis using integrin-α6, GFR-α1, CD90/Thy1 and immunocytochemistry using GFR-α1 antibody showed similar gene expression results. Therefore our results show that iPSC has the potential to differentiate into germ cells and suggest that a protocol optimizing germ cell induction from iPSC should be developed because of their potential usefulness in clinical applications requiring patient-specific cells.

Abstract

In the present study, embryoid bodies (EBs) obtained from induced pluripotent stem cells (iPSCs) were induced to differentiate into germ lineage cells by treatment with bone morphogenetic protein 4 (BMP4) and retinoic acid (RA). The results were compared to the results for embryonic stem cells (ESCs) and multipotent spermatogonial stem cells (mSSCs) and quantified using immunocytochemical analysis of germ cell-specific markers (integrin-α6, GFR-α1, CD90/ Thy1), fluorescence activating cell sorting (FACS), and real time-RT-PCR. We show that the highest levels of germ cell marker-expressing cells were obtained from groups treated with 10 ng/㎖ BMP4 or 0.01 μM RA. In the BMP4-treated group, GFR-α1 and CD90/Thy-1 were highly expressed in the EBs of iPSCs and ESCs compared to EBs of mSSCs. The expression of Nanog was much lower in iPSCs compared to ESCs and mSSCs. In the RA treated group, the level of GFR-α1 and CD90/Thy-1 expression in the EBs of mSSCs was much higher than the levels found in the EBs of iPSCs and similar to the levels found in the EBs of ESCs. FACS analysis using integrin-α6, GFR-α1, CD90/Thy1 and immunocytochemistry using GFR-α1 antibody showed similar gene expression results. Therefore our results show that iPSC has the potential to differentiate into germ cells and suggest that a protocol optimizing germ cell induction from iPSC should be developed because of their potential usefulness in clinical applications requiring patient-specific cells.

참고문헌 (39)

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