Unsicker K, Stahnke G, Müller T H
Department of Anatomy and Cell Biology, University of Marburg, Federal Republic of Germany.
Neurochem Res. 1987 Nov;12(11):995-1003. doi: 10.1007/BF00970928.
Adult bovine and young rat chromaffin cells cultured in serum-free medium were examined for their survival and differentiation following exposure to various additives, trophic agents and conditioned media. Adrenal chromaffin cells dissociated from 8 day old rats were maintained by dexamethasone, NGF and CNTF or without any additives in an N1-supplemented medium in similar numbers as in serum-containing medium for up to 6 days. Neuritic growth elicited by NGF or CNTF was enhanced in the absence of serum. Medium conditioned by purified bovine chromaffin cells improved cell survival and caused neurite outgrowth in a dose-dependent manner. The activiti(es) was sensitive to heat and trypsin and not blocked by the addition of anti-NGF antibodies. Bovine chromaffin cell survival was reduced by 30% when cells were maintained for one week in the absence as compared to the presence of serum. Addition of insulin, the N1 supplement, dexamethasone or dbcAMP single or in combinations improved the survival to different extents. A combination of insulin (5 micrograms/ml) and dexamethasone (5 X 10(-6) M) proved to be optimal in this respect. However, these supplements failed to restore the cellular catecholamine, noradrenaline and adrenaline contents to levels seen in the presence of serum. This was also true for a chromaffin cell-conditioned medium, which improved survival without elevating the catecholamine contents. Conditioned medium, however, partly restored a more physiological adrenaline-noradrenaline-ratio.
在无血清培养基中培养的成年牛和幼年大鼠嗜铬细胞，在暴露于各种添加剂、营养因子和条件培养基后，对其存活和分化情况进行了检测。从8日龄大鼠分离的肾上腺嗜铬细胞，在补充了N1的培养基中，通过地塞米松、神经生长因子（NGF）和睫状神经营养因子（CNTF）维持培养，或者不添加任何添加剂，其数量与在含血清培养基中相似，可维持长达6天。在无血清的情况下，NGF或CNTF引发的神经突生长增强。纯化的牛嗜铬细胞条件培养基可提高细胞存活率，并以剂量依赖的方式引起神经突生长。该活性对热和胰蛋白酶敏感，添加抗NGF抗体不能阻断。与存在血清相比，当细胞在无血清条件下维持一周时，牛嗜铬细胞存活率降低30%。单独或联合添加胰岛素、N1补充剂、地塞米松或二丁酰环磷腺苷（dbcAMP）在不同程度上提高了存活率。在这方面，胰岛素（5微克/毫升）和地塞米松（5×10⁻⁶摩尔/升）的组合被证明是最佳的。然而，这些补充剂未能将细胞儿茶酚胺、去甲肾上腺素和肾上腺素含量恢复到血清存在时的水平。嗜铬细胞条件培养基也是如此，它提高了存活率，但没有提高儿茶酚胺含量。然而，条件培养基部分恢复了更接近生理状态的肾上腺素-去甲肾上腺素比值。
