무 지지세포 (Feeder-Free) 환경에서 히스톤 탈아세틸효소 저해제들을 이용한 인간배아줄기세포와 유도만능줄기세포의 배양방법 (The Feeder-Free Culture of Human Embryonic Stem Cells and Induced Pluripotent Stem Cells in the Presence of Histone Deacetylase Inhibitors)

서지정보

· 발행기관 : 한국조직공학과 재생의학회
· 수록지 정보 : 조직공학과 재생의학 / 9권 / 1호 / 55 ~ 64페이지
· 저자명 : 김형택, 이강인, 황동연

초록

Human pluripotent stem cells (hPSCs) such as human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) retain the capacity to self-renew and differentiate into cells of all three germ layers of the body. These unique properties of hiPSCs make them an ideal cell source for cell-replacement therapies to treat many incurable diseases.
Conventionally, hESCs have been cultured on feeder cells such as mouse embryonic fibroblasts (MEFs) which often hamper successful genetic modification or screening of small molecules. In the previous study, we reported that sodium butyrate, a histone deacetylase inhibitor (HDACi), can replace basic fibroblast growth factor in culture of hESCs using MEFs as feeder cells. However, the effects of HDACis in a feeder-free culture condition were yet to be investigated. Here we set out to explore whether various HDACIs played any role in culture of hPSCs without differentiation. We found that the three commonly used HDACis, sodium butyrate, valproic acid, and trichostatin A supported undifferentiated growth of hESCs and hiPSCs efficiently. In addition, the HDACi-containing culture media also supported stable cell attachment to the bottom of vitronectin-coated dishes during each passage. The expanded hPSCs were shown to express undifferentiation markers at a significant level, while the expression of three germ layer markers was very low. Karyotyping analysis revealed no abnormal change in the chromosomes after culture of hPSCs in these HDACi-containing media. Taken together, our results suggested that HDACis can be useful to design media that can support undifferentiated growth of hPSCs in the absence of feeder cells.

영어초록

Human pluripotent stem cells (hPSCs) such as human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) retain the capacity to self-renew and differentiate into cells of all three germ layers of the body. These unique properties of hiPSCs make them an ideal cell source for cell-replacement therapies to treat many incurable diseases.
Conventionally, hESCs have been cultured on feeder cells such as mouse embryonic fibroblasts (MEFs) which often hamper successful genetic modification or screening of small molecules. In the previous study, we reported that sodium butyrate, a histone deacetylase inhibitor (HDACi), can replace basic fibroblast growth factor in culture of hESCs using MEFs as feeder cells. However, the effects of HDACis in a feeder-free culture condition were yet to be investigated. Here we set out to explore whether various HDACIs played any role in culture of hPSCs without differentiation. We found that the three commonly used HDACis, sodium butyrate, valproic acid, and trichostatin A supported undifferentiated growth of hESCs and hiPSCs efficiently. In addition, the HDACi-containing culture media also supported stable cell attachment to the bottom of vitronectin-coated dishes during each passage. The expanded hPSCs were shown to express undifferentiation markers at a significant level, while the expression of three germ layer markers was very low. Karyotyping analysis revealed no abnormal change in the chromosomes after culture of hPSCs in these HDACi-containing media. Taken together, our results suggested that HDACis can be useful to design media that can support undifferentiated growth of hPSCs in the absence of feeder cells.

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