Gaustad Kristine G, Boquest Andrew C, Anderson Brent E, Gerdes A Martin, Collas Philippe
Institute of Medical Biochemistry, University of Oslo, Oslo, Norway.
Biochem Biophys Res Commun. 2004 Feb 6;314(2):420-7. doi: 10.1016/j.bbrc.2003.12.109.
We report the differentiation of human adipose tissue stem cells (ATSCs) to take on cardiomyocyte properties following transient exposure to a rat cardiomyocyte extract. Reversibly permeabilized ATSCs were incubated for 1h in a nuclear and cytoplasmic extract of rat cardiomyocytes, resealed with CaCl(2), and cultured. Three weeks after exposure to extract, ATSCs expressed several cardiomyocyte markers including sarcomeric alpha-actinin, desmin, and cardiac troponin I, and displayed targeted expression of the gap junction protein connexin 43. Formation of binucleated and striated cells, and spontaneous beating in culture were also observed. A low proportion of intact ATSCs exposed to the extract also showed signs of alpha-actinin and connexin 43 expression. Additional evidence of differentiation was provided by induction of expression of nuclear lamin A/C, a marker of terminally differentiated cells, and a remarkable increase in cell cycle length. Together with our previous data, this study suggests that alteration of cell fate using cellular extracts may be applied to multiple cell types. Cell extracts may also prove useful for investigating the molecular mechanisms of stem cell differentiation.
我们报告了人类脂肪组织干细胞(ATSCs)在短暂暴露于大鼠心肌细胞提取物后呈现心肌细胞特性的分化情况。将可逆性通透化的ATSCs在大鼠心肌细胞核和细胞质提取物中孵育1小时,用氯化钙重新封闭后进行培养。暴露于提取物三周后,ATSCs表达了几种心肌细胞标志物,包括肌节α-肌动蛋白、结蛋白和心肌肌钙蛋白I,并显示出缝隙连接蛋白连接蛋白43的靶向表达。还观察到双核和横纹肌细胞的形成以及培养物中的自发搏动。暴露于提取物的少量完整ATSCs也显示出α-肌动蛋白和连接蛋白43表达的迹象。终末分化细胞标志物核纤层蛋白A/C表达的诱导以及细胞周期长度的显著增加提供了进一步的分化证据。与我们之前的数据一起,这项研究表明使用细胞提取物改变细胞命运可能适用于多种细胞类型。细胞提取物也可能被证明对研究干细胞分化的分子机制有用。