Cheng K F, Her W Y, Liu T S, Chen S C, Liu K M
Department of Anatomy, Kaohsiung Medical College, Kaohsiung City, Taiwan, Republic of China.
Gaoxiong Yi Xue Ke Xue Za Zhi. 1995 Jun;11(6):306-14.
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.
文献显示成肌细胞培养方案存在差异，而成肌细胞和成纤维细胞增殖方面的信息较少。因此，本研究的目的是通过比较先前研究中使用的各种培养程序，建立一种谨慎的成肌细胞培养方案，并对不同培养条件下成肌细胞的增殖和融合进行定量。此外，还研究了成肌细胞和成纤维细胞的生长状态。结果表明，理想的成肌细胞培养条件应包括0.25%胰蛋白酶和0.2%IV型胶原酶（1:1）的混合酶、约15 - 20分钟的预铺板时间以及1×10⁵个细胞/ml的接种密度。此外，小鼠样本应为新生小鼠。在含有10%二氧化碳、杜氏改良 Eagle 培养基加15%胎牛血清的培养箱中，成肌细胞具有更好的增殖能力。成肌细胞的倍增时间比成纤维细胞短，且成肌细胞数量在第4天和第5天达到最高。本研究结果对于了解原代培养中成肌细胞和成纤维细胞的生长状态具有重要价值。此外，确定成肌细胞培养良好生长的条件将有助于成肌细胞移植治疗。
