小鼠成肌细胞的原代培养。

K. Cheng, W. Her, T. S. Liu, S. C. Chen, K. Liu
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引用次数: 0

摘要

文献揭示了成肌细胞培养方案的差异,关于成肌细胞和成纤维细胞增殖的信息很少。因此,本研究的目的是通过比较以往研究中使用的各种培养方法,建立一种谨慎的成肌细胞培养方案,并定量测定不同培养条件下成肌细胞的增殖和融合。同时观察成肌细胞和成纤维细胞的生长状况。结果表明,理想的成肌细胞培养要求包括0.25%胰蛋白酶和0.2% IV型胶原酶(1:1)的组合酶,预镀时间约为15-20分钟,播种密度为1 × 10(5)个细胞/ml。此外,小鼠样本应该是新生儿。在10% CO2的培养箱中,加上Dulbecco的MEM和15%的胎牛血清,发现成肌细胞的增殖能力更好。成肌细胞倍增时间短于成纤维细胞,成肌细胞数量在第4天和第5天达到最高。本研究结果对了解成肌细胞和成纤维细胞在原代培养中的生长状况具有重要意义。此外,建立成肌细胞培养良好生长的要求将促进成肌细胞转移治疗。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Primary culture of mouse myoblasts.
The literature has revealed variations in the protocols for myoblast cultures, and little information is available on myoblast and fibroblast proliferation. Therefore, the purposes of this study were to establish a prudent protocol for myoblast cultures by comparing a variety of culturing procedures used in previous research and to quantitate myoblast proliferation and fusion under different culture conditions. In addition, the growth status of myoblasts and fibroblasts was investigated. Results indicate that the requirements for an ideal myoblast culture should include a combined enzyme of 0.25% trypsin and 0.2% collagenase type IV (1:1), a preplating time of approximately 15-20 minutes, and a seeding density at 1 x 10(5) cells/ml. Furthermore, the mouse sample should be those of newborns. A better proliferative capacity of myoblasts was noted in an incubator of 10% CO2, coupled with Dulbecco's MEM plus 15% fetal calf serum. The doubling times of myoblasts were shorter than those of fibroblasts, and myoblast number reached its highest at 4 and 5 days. The findings of this study are valuable in understanding the growth status of myoblasts and fibroblasts in primary cultures. Moreover, the establishment of requirements for a good growth of myoblast cultures will facilitate myoblast transfer therapy.
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