Shapiro D L, Finkelstein J N, Van Diver T
University of Rochester School of Medicine, Department of Pediatrics (Neonatology), New York 14642.
In Vitro Cell Dev Biol. 1989 Nov;25(11):1051-4. doi: 10.1007/BF02624140.
Lung alveolar epithelial cells have been studied in a variety of laboratory animal models, and studies of human alveolar epithelial cells are important for comparison to information obtained from animal studies. Autopsy material is a source of human cells for study. Studies of human autopsy material revealed variables that negatively affected the yield of viable cells. For specimens from adults, these included death greater than 12 h before cell isolation, obvious severe lung fibrosis, longstanding metabolic disorders, and lung congestion indicated by weight of the right middle lobe greater than 150 g. Samples from children yielded significant numbers of viable cells up to 18 h after death. For 17 specimens that conformed to the above criteria, approximately 8.5 x 10(6) alveolar cells were obtained per gram of tissue (tissue weights ranged from 30 to 108 g) using a procedure involving instillation of proteases into the airways. The cells could be further fractionated, and 10 to 15% of the mixed cells obtained were type II pneumocytes. Analysis of NADPH cytochrome-c-reductase distribution in subcellular fractions provided evidence that the cells obtained were intact. Phospholipid enzyme activities and synthetic activity were within the ranges previously found in laboratory studies of freshly obtained animal lungs. These results suggest that significant numbers of viable and functional human lung cells, including type II pneumocytes, can be obtained from autopsy material.
人们已在多种实验动物模型中对肺泡上皮细胞展开研究,而对人类肺泡上皮细胞的研究对于与从动物研究中获取的信息进行比较至关重要。尸检材料是用于研究的人类细胞来源。对人类尸检材料的研究揭示了一些对活细胞产量产生负面影响的变量。对于成人标本,这些变量包括细胞分离前死亡超过12小时、明显的严重肺纤维化、长期的代谢紊乱以及右中叶重量大于150克所表明的肺充血。儿童样本在死后18小时内可产生大量活细胞。对于符合上述标准的17个标本,通过向气道内滴注蛋白酶的方法,每克组织(组织重量范围为30至108克)可获得约8.5×10⁶个肺泡细胞。这些细胞可进一步分离,所获得的混合细胞中有10%至15%为II型肺细胞。亚细胞组分中NADPH细胞色素c还原酶分布的分析提供了证据,表明所获得的细胞是完整的。磷脂酶活性和合成活性在先前对新鲜获取的动物肺进行实验室研究中所发现的范围内。这些结果表明,可从尸检材料中获得大量有活力且有功能的人类肺细胞,包括II型肺细胞。