• 作者： 鄭光峰; 莊豔鳳; 何宛怡; 陳世杰; 劉克明
• 作者服務機構： 高雄醫學院附設醫院復建科; 高雄醫學院解剖科
• 中文摘要： 　　本研究利用細胞免疫化學染色及甲苯藍對比染色的方法，針對初級正常肌母細胞培養的個樣條件來評估培養條件的差異，進而得到一個培養常模。且利用這個常模，來比較正常與杜顯型肌肉萎縮症之肌母細胞的生長特性。實驗結果顯示：在正常的初級肌母細胞培養，肌母細胞的培養年齡，以剛出生的鼷鼠其體外培養的增殖情形較好；酵素的選擇以0.25％胰蛋白酵素(trypsin)和0.2％膠原酵素(collagenase)(1：1)混合為佳：而最適當的預植時問(preplating time)大約是15-20分鐘；種植密度(seeding density)應控制在1×10 cells/ml。另外，肌母細胞長在Dulbecco's MEM，加上15％胎牛血清，在10％CO2的培養箱裡，其增殖能力表現較佳。在正常與杜顯型肌母細胞的生長特性方面：正常肌母細胞比杜顯型肌母細胞的倍增時間較短（約24-30小時），而杜顯型肌母細胞則約三十五小時。此外，在同一培養天數肌母細胞融合的百分比而言，杜顯型肌母細胞比正常肌母細胞有較早融合的傾向。本研究提供各種不同之培養條件，肌母細胞肌生的變化情形，並從中建立一個培養常模後，可作未來肌母細胞移植療法的應用；在另一方面，也了解到正常與杜顯型肌母細胞的生長特性，並窺知兩者肌母細胞的增殖與融合的差異。
• 英文摘要： This study employed immunocytochemistry and toluidine blue counterstaining to compare differ-ent procedures utilized in primary myoblast cultures, from which an optimal culture model for normalmyoblasts could be derived. The growth characteristics of normal and dystrophic myoblasts were alsoinvestigated by means of this model. Results indicate that the requirements for an ideal myoblast cul-ture should include a combined enzyme of 0.25% trypsin and 0.2% collagenase (type IV) (1 : 1), apreplating time of approximately 15-20 minutes, and a seeding density of 1 x 10 cells/ml. Furthermore,the mouse samples should be newborn mice. A better proliferative capacity of myoblasts was noted inan incubator with 10% CO2 coupled with Dulbecco's MEM plus 15% fetal calf serum. With regard tothe growth characteristics of normal and dystrophic myoblasts, the doubling time of normal myoblastswas shorter than that of dystrophic myoblasts. In terms of the fusion percentage of myoblasts, dystro-phic myoblasts tended to fuse earlier than normal ones, especially after 5 days in culture. The findingsof this study are valuable in understanding the myogenesis of myoblasts under different culture condi-tions. The establishment of requirements for good growth of myoblast cultures will facilitate myoblasttransfer therapy. Finally, the growth characteristics of normal and dystrophic myoblasts as well as vari-ances in the proliferation and differentiation of these two types of cells are clarified.
• 中文關鍵字： myoblast culture; myogenesis; Duchenne muscular dystrophy.
• 英文關鍵字： --