METHODS: Human embryonic stem cells (hESC) differentiated to cardiomyocytes (CM) using a p38 MAPK inhibitor (SB203580) based serum-free medium (SB media). Nutrient supplements known to increase cell viability were added

to SB medium. The ability of these supplements to improve cardiomyogenesis was evaluated by measurements of cell viability, total cell count, and the expression of cardiac markers via flow cytometry. Tariquidar 价格 An improved medium containing Soy hydrolysate (HySoy) and bovine serum albumin (BSA) (SupSB media) was developed and tested on 2 additional cell lines (H1 and Siu-hiPSC). Characterization of the cardiomyocytes was done by immunohistochemistry, electrophysiology and quantitative real-time reverse transcriptionpolymerase chain reaction. RESULTS: hESC cell line, HES-3, differentiating in SB medium for 16 d resulted in a cardiomyocyte yield of 0.07 ± 0.03 CM/hESC. A new medium (SupSB media) was developed with the addition of HySoy and BSA to SB medium. This medium resulted in 2.6 fold increase in cardiomyocyte yield (0.21

± 0.08 CM/hESC). The robustness of SupSB medium was further demonstrated VE-821购买 using two additional pluripotent cell lines (H1, hESC and Siu1, hiPSC), showing a 15 and 9 fold increase in cardiomyocyte yield respectively. The age (passage number) of the pluripotent cells did not affect the cardiomyocyte yields. Embryoid body (EB) cardiomyocytes formed in SupSB medium expressed canonical cardiac markers (sarcomeric α-actinin, myosin heavy chain and troponin-T) and demonstrated all three major phenotypes: nodal-, atrial- and ventricular-like. Electrophysiological characteristics (maximum diastolic potentials and action potential durations) of cardiomyocytes derived from SB and SupSB media were similar. CONCLUSION:

The nutrient supplementation (HySoy and BSA) leads to increase in cell viability, cell yield and cardiac marker expression during cardiomyocyte Doxorubicin化学结构 differentiation, translating to an overall increase in cardiomyocyte yield.
目的探讨乳凝集素通过ERK、p38 MAPK信号通路对未成熟树突状细胞(iDCs)分泌白介素(IL)-12和IL-10的影响。方法脐带血中分离得到单核细胞,加入重组人粒细胞巨噬细胞集落刺激因子(50 ng/mL)和重组人白介素-4(10 ng/mL),同时按照不同组合加入乳凝集素(10μg/mL)和信号通路阻滞剂,诱导分化为iDCs。分组如下:①对照组;②乳凝集素组;③ERK通路阻滞组;④ERK通路阻滞剂+乳凝集素组;⑤p38MAPK通路阻滞组;⑥p38MAPK通路阻滞剂+乳凝集素组;⑦JNK通路阻滞组;⑧JNK通路阻滞剂+乳凝集素组。采用Western blotting检测ERK、JNK、p38MAPK蛋白磷酸化水平,ELISA法检测iDCs分泌IL-12和IL-10的变化。结果与对照组比较,乳凝集素组ERK蛋白磷酸化水平升高(P0.05),IL-12和IL-10的分泌量显著降低(P<0.05)。ERK通路阻滞组和ERK通路阻滞剂+乳凝集素组中ERK蛋白磷酸化水平几乎为零;ERK阻滞剂+乳凝集素组IL-12和IL-10的分泌量显著高于乳凝集素组(P<0.