Andersson A, Christensen N, Groth C G, Hellerström C, Petersson B, Sandler S
Transplantation. 1984 May;37(5):499-503. doi: 10.1097/00007890-198405000-00015.
Human fetal pancreatic explants were maintained in organ culture for up to 12 days in medium TC 199 or RPMI 1640 supplemented with 20% fetal calf serum. The explants were cultured at their air-liquid interface on a Millipore filter supported by surgical gel foam (ALI technique). The insulin accumulation in the culture medium decreased considerably when explants were maintained for 6 days in RPMI 1640. No corresponding decline was observed with medium TC 199. Four out of 9 RPMI 1640-cultured and 8 out of 11 TC 199-cultured explants responded with an increased insulin release when challenged with high glucose-plus-theophylline after one week of culture. With these explants there was a statistically significant correlation between their rates of respiration and insulin secretion. Furthermore, significant correlations were found between the fetal crown-heel length on the one hand--and the insulin content, insulin response, and oxygen uptake on the other--of the TC 199 cultured explants. There was no apparent change in the rate of insulin accumulation by addition of high glucose or amino acids to the TC 199 medium. It is concluded that oxygen uptake measurements, requiring only minute amounts of tissue, are useful for assessing the viability of human fetal pancreatic explants, and that the ALI technique for maintaining human fetal pancreas in culture offers no obvious advantage over that of the submersion technique. There is also evidence that suggests a better functional preservation in culture medium TC 199 than in RPMI 1640.
将人胎儿胰腺外植体在添加20%胎牛血清的TC 199培养基或RPMI 1640培养基中进行器官培养长达12天。外植体在气液界面的密理博滤膜上培养,滤膜由外科手术用凝胶泡沫支撑(气液界面技术)。当外植体在RPMI 1640中培养6天时,培养基中的胰岛素积累显著减少。而在TC 199培养基中未观察到相应下降。培养一周后,用高糖加茶碱刺激时,9个在RPMI 1640中培养的外植体中有4个、11个在TC 199中培养的外植体中有8个胰岛素释放增加。对于这些外植体,其呼吸速率与胰岛素分泌之间存在统计学上的显著相关性。此外,在TC 199培养的外植体中,一方面胎儿顶臀长与另一方面胰岛素含量、胰岛素反应和氧摄取之间也发现了显著相关性。向TC 199培养基中添加高糖或氨基酸后,胰岛素积累速率没有明显变化。结论是,仅需极少量组织的氧摄取测量对于评估人胎儿胰腺外植体的活力是有用的,并且在培养中维持人胎儿胰腺的气液界面技术相对于浸没技术没有明显优势。也有证据表明,TC 199培养基比RPMI 1640培养基在培养中能更好地保持功能。