Malaver-Ortega Luis F, Sumer Huseyin, Liu Jun, Verma Paul J
CSIRO Health & Biosecurity, East Geelong, VIC 3219, Australia; Monash Institute for Medical Research, Monash University, Clayton, VIC 3168, Australia.
Faculty of Science, Engineering and Technology, Swinburne University of Technology, Hawthorn, VIC 3122, Australia.
Stem Cells Int. 2016;2016:5127984. doi: 10.1155/2016/5127984. Epub 2016 Jan 6.
Pluripotent stem cells (PSCs) fall in two states, one highly undifferentiated, the naïve state, and the primed state, characterized by the inability to contribute to germinal lineage. Several reports have demonstrated that these states can be modified by changes to the cell culture conditions. With the advent of nuclear reprogramming, bovine induced pluripotent stem cells (biPSCs) have been generated. These cells represent examples of a transient-intermediate state of pluripotency with remarkable characteristics and biotechnological potential. Herein, we generated and characterized biPSC. Next, we evaluated different culture conditions for the ability to affect the expression of the set of core pluripotent transcription factors in biPSC. It was found that the use of 6-bromoindirubin-3-oxime and Sc1 inhibitors alone or in combination with 5-AzaC induced significantly higher levels of expression of endogenous REX1, OCT4, NANOG, and SOX2. Furthermore, LIF increased the levels of expression of OCT4 and REX1, compared with those cultured with LIF + bFGF. By contrast, bFGF decreased the levels of expression for both REX1 and OCT4. These results demonstrate that the biPSC gene expression profile is malleable by modification of the cell culture conditions well after nuclear reprogramming, and the culture conditions may determine their differentiation potential.
多能干细胞（PSCs）处于两种状态，一种是高度未分化的原始状态，另一种是始发态，其特征是无法分化为生殖系细胞。多项报告表明，这些状态可通过改变细胞培养条件来改变。随着核重编程技术的出现，已成功诱导出牛诱导多能干细胞（biPSCs）。这些细胞代表了一种具有显著特征和生物技术潜力的多能性瞬时中间状态。在此，我们成功诱导并鉴定了biPSCs。接下来，我们评估了不同培养条件对biPSCs中一组核心多能转录因子表达的影响。结果发现，单独使用6-溴靛玉红-3-肟和Sc1抑制剂或与5-氮杂胞苷联合使用，可显著提高内源性REX1、OCT4、NANOG和SOX2的表达水平。此外，与在LIF + bFGF条件下培养的细胞相比，LIF可提高OCT4和REX1的表达水平。相反，bFGF会降低REX1和OCT4的表达水平。这些结果表明，在核重编程后，通过改变细胞培养条件可以改变biPSCs的基因表达谱，并且培养条件可能决定它们的分化潜能。
