Dunger A, Besch W, Hahn H J
Institut für Diabetes Gerhardt Katsch, Karlsburg, Germany.
Horm Metab Res. 1991 May;23(5):201-4. doi: 10.1055/s-2007-1003653.
Pancreatic islets obtained from newborn Lewis rats were kept free-floating in TCM 199 either in the presence of 50% human amniotic fluid (HAF), 50% Hank's balanced salt solution (HBSS) or 10% fetal calf serum (FCS) for up to 14 days. The culture, in the presence of HAF, resulted in a good islet viability as demonstrated by islet number, islet insulin content and insulin release into the medium. Contradictory results were obtained when HBSS was used as the medium supplement. Moreover, the islets cultured in HAF-supplemented medium are characterized by a marked replicatory activity as reflected by the incorporation of labeled thymidine into islet DNA and gradual increase in DNA content with the progression of culture time. Even if compared to DNA synthesis of islets cultured under so called standard culture conditions as e.g. supplementation of 10% FCS to the medium, 50% HAF but not yet 10% HAF was found to stimulate the islet replication twofold already after 4 days of culture. It was also demonstrated that FCS has no additional effect on HAF-stimulated DNA synthesis. These findings support the view that HAF-enriched culture medium may have a use in supporting the long-term survival of well preserved endocrine pancreatic tissue.
从新生Lewis大鼠获取的胰岛,在含有50%人羊水(HAF)、50%汉克平衡盐溶液(HBSS)或10%胎牛血清(FCS)的TCM 199中自由漂浮培养长达14天。在HAF存在的情况下进行培养,通过胰岛数量、胰岛胰岛素含量以及胰岛素释放到培养基中的情况表明,胰岛具有良好的活力。当使用HBSS作为培养基补充物时,得到了相互矛盾的结果。此外,在添加HAF的培养基中培养的胰岛具有显著的复制活性,这通过将标记的胸苷掺入胰岛DNA以及随着培养时间的延长DNA含量逐渐增加得以体现。即使与在所谓标准培养条件下(例如向培养基中添加10% FCS)培养的胰岛的DNA合成相比,发现50% HAF而非10% HAF在培养4天后就已能使胰岛复制增加两倍。还证明FCS对HAF刺激的DNA合成没有额外影响。这些发现支持了这样一种观点,即富含HAF的培养基可能有助于维持保存良好的内分泌胰腺组织的长期存活。