大鼠脑皮质星形胶质细胞的限制性细胞培养

介绍一种新的脑组织星形胶质细胞培养方法即限制性细胞培养（constraint cell culture）。常规分离纯化星形胶质细胞，将其低密度种植，维持在添中低量血清的化学成分限定的培养基中培养，并在长时期内不给予更换或补加培养液。利用波形蛋白(vimentin）和胶质纤维酸性蛋白（glial fibrary acidic protein）抗体的免疫荧光染色法鉴定观察不同培养时期的星形胶质细胞及其形态学变化。结果发现星形胶质细胞在最初的5天之内有一定程度的增殖，未出现过度增殖导致的细胞相互融合现象；接下来的3－5天内细胞形态明显分化，星形胶质细胞突起细长、胞体明显缩小、形态多样，最后细胞突起之间相互连接形成星形胶质细胞网络，并在相当长的时间内保持不变。实验结果显示在限制细胞种植密度和限制给予培养液的培养条件下星形质细胞的体外形态发育与在体的情形基本一致。提示该细胞培养方法可能有助于研究中枢神经系统中星形胶质细胞的生理功能。

A RESTRAINT CELL CULTURE OF ASTROCYTES FROM RAT CEREBRAL TISSUE

A novel method termed constraint cell culture for astrocytes harvested from rat cerebral tissue has been developed. The astrocytes were first purified as discribed previously, and the cells were replanted at low density and maintained for a long peroid in a chemical difinitative medium with little serum added. Vimentin and glial fibrary acidic protein immunofluorescence staining was used to investigate the morphological changes of astrocytes at different time in culture. It was found that astrocytes could proliferate in certain degree during the first 5 days but not overgrow to confluence, and in the following 3 to 5 days differentiation of astrocytes produced a variety of morphologically distinct cells with many long processes and small cell bodies. Finally formation of astrocytic network occured through process connections, and this kind of network could maintain over a relatively broad period. The results indicated that astrocytes in the constraint cell culture had the ability to undergo a morphological development similar to that taking place in vivo. so this method could be helpful to study the physiological functions of astrocytes in the central nervous system.;