Isolation of highly pure alveolar epithelial type I and type II cells from rat lungs
Isolation of highly pure alveolar epithelial type I and type II cells from rat lungs
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DOI：
10.1038/labinvest.3700095
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发表时间：
2004-06-01
影响因子：
5
通讯作者：
Liu, L
中科院分区：
文献类型：
--
作者：
Chen, JW;Chen, ZM;Liu, L
目前还没有理想的细胞系可用于肺泡上皮I型和II型细胞（AEC I和II）。目前分离AEC I和II的方法纯度有限。在这里，我们报告了改进的和可重复的方法，用于从大鼠肺中分离高纯度的AEC I和II。使用不同浓度的弹性蛋白酶消化，AEC I和II从肺组织中释放出来。通过大鼠IgG“淘洗”和抗大鼠白细胞共同抗原抗体去除巨噬细胞和白细胞。对于AEC 11分离，使用多克隆兔抗T1 α（AEC I顶端膜蛋白）抗体去除AEC I污染。对于AEC I分离，使用多克隆抗T1 α抗体的阳性免疫磁性选择。AEC I和II的纯度分别为91 +/- 4和97 +/-1%。AEC I和AEC II的每只大鼠产量分别为2 x 10（6）和33 x 10（6）。这些细胞制备物的活力大于96%。AEC II分离方案也适用于从高氧损伤和恢复的肺中获得纯AEC II（93-95%）。纯化的AEC I和II可用于基因表达谱和功能研究。它也为肺生物学领域提供了重要工具。
There are no ideal cell lines available for alveolar epithelial type I and II cells (AEC I and II) at the present time. The current methods for isolating AEC I and II give limited purities. Here, we reported improved and reproducible methods for the isolation of highly pure AEC I and II from rat lungs. AEC I and II were released from lung tissues using different concentrations of elastase digestion. Macrophages and leukocytes were removed by rat IgG 'panning' and anti-rat leukocyte common antigen antibodies. For AEC 11 isolation, polyclonal rabbit anti-T1alpha (an AEC I apical membrane protein) antibodies were used to remove AEC I contamination. For AEC I isolation, positive immunomagnetic selection by polyclonal anti-T1alpha antibodies was used. The purities of AEC I and II were 91 +/- 4 and 97 +/- 1 %, respectively. The yield per rat was similar to 2 x 10(6) for AEC I and similar to 33 x 10(6) for AEC II. The viabilities of these cell preparations were more than 96%. The protocol for AEC II isolation is also suitable to obtain pure AEC II (93-95%) from hyperoxia-injured and recovering lungs. The purified AEC I and II can be used for gene expression profiling and functional studies. It also offers an important tool to the field of lung biology.