METABOLIC PROPERTIES AND ULTRASTRUCTURE OF ALVEOLAR TYPE-II CELLS ISOLATED WITH ELASTASE
METABOLIC PROPERTIES AND ULTRASTRUCTURE OF ALVEOLAR TYPE-II CELLS ISOLATED WITH ELASTASE
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DOI：
10.1016/0005-2760(80)90270-2
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发表时间：
1980-01-01
期刊：
影响因子：
--
通讯作者：
MASON, RJ
中科院分区：
文献类型：
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作者：
DOBBS, LG;GEPPERT, EF;MASON, RJ
使用猪胰腺弹性蛋白酶从大鼠肺中分离II型细胞。II型细胞的产量和纯度优于使用胰蛋白酶的方法获得的。在102个实验中，82 .+-. 23.cntdot.获得106个细胞/大鼠，68 ± 100个。11%（平均值±）SD），其中II型细胞。当在不连续的密度梯度上离心时，这种细胞制备物产生25 ± 0.5%。10. cntdot. 106个细胞/大鼠，80 . ±.其中13%为II型细胞（n = 102）。在密度梯度离心后，细胞可以通过离心淘析（94 ± 0.01）进一步纯化。3%II型细胞，n = 22）或原代培养物中的粘附（94 . ±. 2%的II型细胞，n = 34）。用弹性蛋白酶分离的II型细胞与用胰蛋白酶从大鼠分离的II型细胞在形态学和生物化学上相似。几个不同的标准：超微结构，排除活体染料，缺乏刺激的O2消耗的外源性琥珀酸钠和线性速率的氧化[1- 14 C]棕榈酸和[1- 14 C]乙酸纳入脂肪酸的细胞制剂似乎健康。II型细胞消耗75 . ±. 20 nmol O2/106细胞/h，氧化[1- 14 C]棕榈酸的速率为0.4 nmol/106细胞/h，[1- 14 C]乙酸酯掺入脂肪酸的速率为7.5 nmol/106细胞/h。
Porcine pancreatic elastase was used to isolate type II cells from the lungs of rats. Yield and purity of the type II cells were better than that obtained by methods using trypsin. In 102 experiments 82 .+-. 23 .cntdot. 106 cells/rat were obtained, 68 .+-. 11% (mean .+-. SD) of which were type II cells. This preparation of cells, when centrifuged over a discontinuous density gradient, yielded 25 .+-. 10 .cntdot. 106 cells/rat, 80 .+-. 13% of which were type II cells (n = 102). The cells, after density gradient centrifugation, could be further purified by centrifugal elutriation (94 .+-. 3% type II cells, n = 22) or adherence in primary culture (94 .+-. 2% type II cells, n = 34). Type II cells isolated with elastase are similar morphologically and biochemically to type II cells isolated from rats with trypsin. The preparations of cells appeared healthy by several different criteria: ultrastructure, exclusion of vital dye, lack of stimulation of O2 consumption by exogenous sodium succinate and linear rates of oxidation of [1-14C]palmitic acid and of incorporation of [1-14C]acetate into fatty acids. Type II cells consumed 75 .+-. 20 nmol O2/106 cell per h, oxidized [1-14C]palmitic acid at a rate of 0.4 nmol/106 cells per h and incorporated [1-14C]acetate into fatty acids at a rate of 7.5 nmol/106 cells per h.