Oomen L C, Ten Have-Opbroek A A, Hageman P C, Oudshoorn-Snoek M, Egberts J, van der Valk M A, Calafat J, Demant P
Department of Molecular Genetics, The Netherlands Cancer Institute, Antoni van Leeuwenhoek Huis, Amsterdam.
Am J Respir Cell Mol Biol. 1990 Oct;3(4):325-39. doi: 10.1165/ajrcmb/3.4.325.
Alveolar type II cells were isolated from fetal mouse lung by differential adherence and obtained in monolayer culture. Cultures display a high degree of purity as shown by histochemical and immunocytochemical staining procedures. Seventy-five percent of cells stained positive with specific anti-lavage serum mouse (SALS-M), an antiserum specific for (pre)alveolar type II cells of the mouse, and osmiophilic bodies were present in 82% of cells. These and other characteristics of type II cells in culture correspond to those of alveolar type II cells in fetal mouse lung. The pattern of reactivity of these cells with various anti-cytokeratin antibodies is described, and we show that, in contrast to rat type II cells, they do not exhibit alkaline phosphatase activity. Identity of the type II cell cultures was shown by their specific phospholipid composition and surfactant protein A (SP-A) content. The fetal alveolar type II cells in culture were found to synthesize and express class I but not class II major histocompatibility complex (MHC) antigens. The possibility to culture fetal alveolar type II cells of the mouse and the availability of genetically well-defined inbred and transgenic mouse strains opens ways to study the genetics of type II cell differentiation and function. Also, the in vitro availability of alveolar type II cells, the progenitor cells of mouse lung tumors, will enable us to study in vitro several of the processes involved in lung tumorigenesis in the mouse.
通过差异贴壁法从小鼠胎儿肺中分离出II型肺泡细胞,并进行单层培养。如组织化学和免疫细胞化学染色程序所示,培养物显示出高度的纯度。75%的细胞用特异性抗小鼠灌洗血清(SALS-M)染色呈阳性,该抗血清对小鼠(前)II型肺泡细胞具有特异性,并且82%的细胞中存在嗜锇性小体。培养的II型细胞的这些及其他特征与小鼠胎儿肺中的II型肺泡细胞的特征相符。描述了这些细胞与各种抗细胞角蛋白抗体的反应模式,并且我们表明,与大鼠II型细胞不同,它们不表现出碱性磷酸酶活性。II型细胞培养物的特性通过其特定的磷脂组成和表面活性蛋白A(SP-A)含量得以体现。发现培养的胎儿II型肺泡细胞可合成并表达I类而非II类主要组织相容性复合体(MHC)抗原。培养小鼠胎儿II型肺泡细胞的可能性以及基因定义明确的近交系和转基因小鼠品系的可用性为研究II型细胞分化和功能的遗传学开辟了道路。此外,作为小鼠肺肿瘤祖细胞的II型肺泡细胞在体外的可得性将使我们能够在体外研究小鼠肺肿瘤发生过程中涉及的若干过程。