THIAZOVIVIN

Thiazovivin是一种新型ROCK抑制剂，IC50为0.5 μM，在单细胞分离后，促进人胚胎干细胞（hESC）的存活。 Although displaying little impact on cell proliferation, Thiazovivin treatment significantly enhances the survival of human embryonic stem cells (hESCs) after enzymatic dissociation more than 30-fold, while homogenously maintaining pluripotency with the characteristic colony morphology, expression of typical pluripotency markers such as alkaline phosphatase (ALP), and normal karyotype. Dissociated hESCs treated with Thiazovivin display dramatically increased adhesion to matrigel- or laminin-coated plates but not to gelatin-coated plates within a few hours. Thiazovivin treatment increases cell-ECM adhesion-mediated β1 integrin activity, which synergizes with growth factors to promote cell survival. In addition to activating integrin, Thiazovivin but not Tyrintegin (Ptn) protects hESCs from death in the absence of ECM in suspension through E-cadherin-mediated cell-cell interaction. Thiazovivin treatment potently inhibits endocytosis of E-cadherin, consequently stabilizing E-cadherin on the cell surface and leading to reestablishment of cell-cell interaction, which is essential for hESC survival in ECM-free conditions. Thiazovivin but not Tyrintegin (Ptn) at 2 μM inhibits Rho-associated kinase (ROCK) activity and protects hESCs at a similar level as the widely used selective ROCK inhibitor Y-27632 at 10 μM, suggesting that Rho-ROCK signaling regulates cell-ECM and cell-cell adhesion. Thiazovivin at 1 μM increases the reprogramming efficiency of CB mononuclear cells to induced pluripotent stem cells (iPSCs) by more than 10 times. Thiazovivin是一种新型ROCK抑制剂，无细胞试验中IC50为0.5 μM，在单细胞分离后，促进人胚胎干细胞（hESC）的存活。

Target	Value
ROCK (Cell-free assay)	~0.5 μM

尽管对细胞增殖影响较小，Thiazovivin治疗显著增强人胚胎干细胞(hESCs)的存活，使其被酶解后的存活增强30多倍，同时均匀维持特有集落形态的多潜能，和典型多潜能标志物，如碱性磷酸酶(ALP) 的表达，和正常核型。解离的hESCs用Thiazovivin处理几小时内显著增加其对人工基底膜-或层粘连蛋白涂覆板的粘附，而对明胶涂覆的平板没有影响。Thiazovivin治疗增加细胞-ECM粘附介导的β1整合蛋白活性，其与生长因子协同促进细胞存活。除了活化整合蛋白，Thiazovivin，而不是Tyrintegin (Ptn)，也能够通过E-钙黏蛋白介导的细胞-细胞相互作用，保护hESCs在ECM缺乏的悬浮液中免于死亡。Thiazovivin治疗有效抑制E-钙黏蛋白的内吞作用，从而稳定细胞表面的E-钙黏蛋白，并使细胞-细胞相互作用恢复，这对hESC在无ECM环境下的存活是必须的。Thiazovivin，而不是Tyrintegin (Ptn)，在2 μM浓度下抑制Rho相关的激酶(ROCK)活性，并保护hESCs，与广泛使用的选择性ROCK抑制剂Y-27632在10 μM浓度下的作用类似，表明Rho-ROCK信号调节细胞-ECM和细胞-细胞粘附。1 μM Thiazovivin增加CB单核细胞的重组效能，以诱导10倍以上的多功能干细胞(iPSCs)。