Leary J F, Finkelstein J N, Notter R H, Shapiro D L
Cytometry. 1986 Sep;7(5):431-8. doi: 10.1002/cyto.990070507.
Cell populations dissociated from fetal rabbit lungs were analyzed by laser flow cytometry for the presence of type II pneumocytes. These cells are distinguishable by the staining of their lamellar bodies with the fluorescent lipophilic dye, phosphine-3R and by their intensity of low-angle light scatter. Lung cells were obtained by enzymatic dissociation from fetal rabbits at gestational ages of 24 d, 27 d, and from 2-d newborn rabbits. Flow cytometric analysis was sufficiently sensitive to discriminate between fetuses. Quantitative analysis of type II pneumocytes showed that newborn rabbits had a distinct cell subpopulation in a region of low-angle light scatter and phosphine-3R fluorescence intensity similar to that previously reported on type II cells from adult rabbits. By contrast, 24-d gestation rabbits had a negligible type II cell subpopulation. Fetuses of 27 and 30 d gestation showed a slow but progressive increase in the numbers of cells in the type II region. Mathematical analyses of light scatter and fluorescence intensity distributions were used to define statistically significant (P less than .05) boundaries that characterize the development of the type II cell subpopulation in fetal rabbit lung. The methods employed offer new possibilities for quantification of developing lung cell subpopulations of particular interest to the problem of respiratory distress syndrome in human neonates.
通过激光流式细胞术分析从胎兔肺中解离出的细胞群体，以检测II型肺细胞的存在。这些细胞可通过用荧光亲脂性染料磷化氢-3R对其板层小体进行染色以及通过其低角度光散射强度来区分。肺细胞是通过酶解从妊娠24天、27天的胎兔以及出生2天的新生兔中获得的。流式细胞术分析足够灵敏，能够区分不同的胎儿。对II型肺细胞的定量分析表明，新生兔在低角度光散射和磷化氢-3R荧光强度区域有一个独特的细胞亚群，类似于先前报道的成年兔II型细胞。相比之下，妊娠24天的兔的II型细胞亚群数量可以忽略不计。妊娠27天和30天的胎儿在II型区域的细胞数量显示出缓慢但逐渐增加的趋势。利用光散射和荧光强度分布的数学分析来定义具有统计学意义（P小于0.05）的界限，这些界限表征了胎兔肺中II型细胞亚群的发育。所采用的方法为量化发育中的肺细胞亚群提供了新的可能性，这对于人类新生儿呼吸窘迫综合征问题特别有意义。
