Parimary Culture of Mouse Myoblasts

過去培養肌母細胞的方法，仍存有許多的義異性，並且在培養過程，肌母細胞與纖維母細胞的增殖情形，了解也十分有限。因此，本篇研究的目地有二：一、比較過去許多方法的差異後，得到一個理想的培養條件。二、評估肌母細胞在不同條件下，其增殖融合情形，並且從中探討纖維母細胞和肌母細胞的生長情形。結果顯示：肌肉年齡的選取以院出生的小鼠較好；酵素的選擇以0.25%胰蛋白酵素和0.2%膠原酵素(1：1)混合為佳；而最適當的預植時間(pre-plating time)大約是15-20分鐘；種植密度(seeding density)應控制在1*10的5次方 cells/ms。另外，肌母細胞長在Dulbecco’s MEM，加上15%牛胎兒血清，在10% CO2的培養箱裏，其增殖能力表現較好。另外肌母細胞比纖維母細胞的倍增時間還短，約培養在第四~五天，肌母細胞的數目達到最高。本篇可了解到肌母細胞與纖維母細胞的生長情形，並且在得到一個理想的培養條件後，可供應用於作肌母細胞移植療法(myoblast transfer therapy)的基礎。

關鍵字

無資料

並列摘要

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(1) to establish a prudent protocol for myoblast cultures by comparing a variety of culturing procedures used in previous research and (2) 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*105 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 Dulecco’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.

並列關鍵字

myoblast culture ； myogenesis ； fusion

國際替代計量

Parimary Culture of Mouse Myoblasts

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