鼠胚髁突外植体无血清培养模型的建立与评价

Growth and histodifferentiation of the condylar explant of the fetal mouse in serum-free organ culture system

目的体外建立髁突软骨外植体无血清培养模型,为研究髁突软骨发育改建及各种因子对髁突软骨作用提供适宜手段。方法体外解剖分离鼠胚髁突,行外植体培养,通过组织学、免疫组织化学等方法观察髁突外植体在无血清和含血清体外培养系统中的生长和组织分化状况。结果在无血清培养20天后,髁突软骨膜与软骨层保持紧密结合,尽管部分肥大层软骨排列紊乱,阿辛蓝及Ⅱ、X型胶原染色显示培养软骨组织能形成软骨基质。在含血清的对照组中,软骨组织分化层次过程紊乱,纤维层过度增生并与软骨分离,软骨基质减少,软骨部分降解。结论此无血清培养髁突外植体模型中髁突软骨能正常发育,髁突软骨细胞增殖与分化与体内状态接近,此培养系统可用来体外研究髁突发育以及各种环境因子对髁突生长、发育、生理改建的影响。

Objective To establish a model of the condylar explant in serum - free organ culture for study the development and the mechanism of physiological rebuilding and restoration of condylar cartilage, Methods The explant of condylar in the fetal mouse was separated and cultured in vitro and the influence of a serum - supplemented or serum - free medium on its endochondral oesification of condylar cartilage was observed by immunohistochemistry methods. Results After 20 days culture in serum - free Conditions, the original cartilage mass was maintained as one unit, eel[ proliferation was found to differentiate functional chondroblast. Perichondrium was often tightness with the condylar cartilage. Despite the disintegration of a portion of hypertrophic cartilage, the alcian - blue and Ⅱ. Xcollegen immunostaining pattern showed matrix formation continued. In control cultured with 10% fetal bovine serum, the cartilage histodifferentiation was disturbed during the incubation period. Perichondrium often overgrew and separated the condyar cartilage, and matrix formation was decreased or portions of cartilage were degenerated. Conclusion The condylar explant exhibited normal growth and histodifferentiation in serum - free organ culture. It is a good model to study the development of condylar and to analyze influence of many kinds of environmental factors on its growth, development and physiological rebuilding.