Li Yuan-Yuan, Lu Shan-Shan, Xu Ting, Zhang Hong-Qi, Li Hua
State Key Laboratory of Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Chin Med J (Engl). 2015 Jul 20;128(14):1942-7. doi: 10.4103/0366-6999.160560.
This study characterized the cardiac telocyte (TC) population both in vivo and in vitro, and investigated its telomerase activity related to mitosis.
Using transmission electron microscopy and a phase contrast microscope, the typical morphological features of cardiac TCs were observed; by targeting the cell surface proteins CD117 and CD34, CD117 + CD34 + cardiac TCs were sorted via flow cytometry and validated by immunofluorescence based on the primary cell culture. Then the optimized basal nutrient medium for selected population was examined with the cell counting kit 8. Under this conditioned medium, the process of cell division was captured, and the telomerase activity of CD117 + CD34 + cardiac TCs was detected in comparison with bone mesenchymal stem cells (BMSCs), cardiac fibroblasts (CFBs), cardiomyocytes (CMs).
Cardiac TCs projected characteristic telopodes with thin segments (podomers) in alternation with dilation (podoms). In addition, 64% of the primary cultured cardiac TCs were composed of CD117 + CD34 + cardiac TCs; which was verified by immunofluorescence. In a live cell imaging system, CD117 + CD34 + cardiac TCs were observed to enter into cell division in a short time, followed by an significant invagination forming across the middle of the cell body. Using a real-time quantitative telomeric-repeat amplification assay, the telomerase concentration in CD117 + CD34 + cardiac TCs was obviously lower than in BMSCs and CFBs, and significantly higher than in CMs.
Cardiac TCs represent a unique cell population and CD117 + CD34 + cardiac TCs have relative low telomerase activity that differs from BMSCs, CFBs and CMs and thus they might play an important role in maintaining cardiac homeostasis.
本研究对心脏telocyte(TC)群体进行了体内和体外特征分析,并研究了其与有丝分裂相关的端粒酶活性。
使用透射电子显微镜和相差显微镜观察心脏TC的典型形态特征;通过靶向细胞表面蛋白CD117和CD34,利用流式细胞术分选CD117 + CD34 +心脏TC,并基于原代细胞培养通过免疫荧光进行验证。然后使用细胞计数试剂盒8检测所选群体的优化基础营养培养基。在这种条件培养基下,捕捉细胞分裂过程,并与骨髓间充质干细胞(BMSC)、心脏成纤维细胞(CFB)、心肌细胞(CM)比较检测CD117 + CD34 +心脏TC的端粒酶活性。
心脏TC伸出具有薄段(podomers)与扩张(podoms)交替的特征性telopodes。此外,原代培养的心脏TC中有64%由CD117 + CD34 +心脏TC组成;这通过免疫荧光得到验证。在活细胞成像系统中,观察到CD117 + CD34 +心脏TC在短时间内进入细胞分裂,随后在细胞体中部形成明显的内陷。使用实时定量端粒重复序列扩增测定法,CD117 + CD34 +心脏TC中的端粒酶浓度明显低于BMSC和CFB,且显著高于CM。
心脏TC代表一种独特的细胞群体,CD117 + CD34 +心脏TC具有相对较低的端粒酶活性,这与BMSC、CFB和CM不同,因此它们可能在维持心脏内环境稳定中起重要作用。