Growth characteristics, morphology, and phospholipid composition of human type II pulmonary alveolar cells grown in a collagen-free microenvironment.

In Vitro Pub Date : 1984-12-01 DOI:10.1007/BF02619663
G E Milo, G A Ackerman, R L Sanders
{"title":"Growth characteristics, morphology, and phospholipid composition of human type II pulmonary alveolar cells grown in a collagen-free microenvironment.","authors":"G E Milo,&nbsp;G A Ackerman,&nbsp;R L Sanders","doi":"10.1007/BF02619663","DOIUrl":null,"url":null,"abstract":"<p><p>Human lung epithelial cells have been isolated and maintained in pure culture and characterized during their time in culture. Any residual fibroblasts were removed by selective trypsinization within the first 48 h in culture and the residual epithelial cells from the primary culture grew to confluent density. The epithelial cells at Passage 2 or greater were serially subpassaged when cultures reached ca. 80% confluency. This procedure permitted us to conduct biochemical and structural studies of starting materials and subsequent population doublings. Electron microscope evaluation of both initial monolayers and cell suspensions showed cultures to be composed of a single cell type. These cells had microvilli on their free or apical surface. Subsequent population doubling level 1 up to 5 exhibited the same structures. They contained lamellar inclusions, which are typical of Type II alveolar epithelial cells. Fetal lung (age 18 to 20 wk) cell suspensions processed for electron microscopy before culturing showed cells to be undifferentiated, epithelial-like with small microvilli along cell borders, and with desmosomes at cell junctions. Lamellar inclusions were not observed in these cells. Ultrastructural studies of the cultured epithelial cells demonstrated that the lamellar inclusions had a slightly positive reaction when tested for acid phosphatase. Phospholipid analysis of these lung epithelial cells showed a phospholipid composition consistent with that found in surfactant-containing Type II cells. Cultured epithelial cells stained with phosphine 3-R demonstrated a green fluorescent cytoplasm and nucleus with brightly fluorescent yellow-orange perinuclear particles. The preceding characterization of these cells leads us to conclude that they exhibit structural and biochemical features commensurate with Type II epithelial cells from human lung. Moreover, these selection techniques applied to the isolation of human lung Type II cells from the tissue permit us to study the differentiative function of these cells routinely under conditions of growth in vitro.</p>","PeriodicalId":13317,"journal":{"name":"In Vitro","volume":"20 12","pages":"899-911"},"PeriodicalIF":0.0000,"publicationDate":"1984-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02619663","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"In Vitro","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1007/BF02619663","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 1

Abstract

Human lung epithelial cells have been isolated and maintained in pure culture and characterized during their time in culture. Any residual fibroblasts were removed by selective trypsinization within the first 48 h in culture and the residual epithelial cells from the primary culture grew to confluent density. The epithelial cells at Passage 2 or greater were serially subpassaged when cultures reached ca. 80% confluency. This procedure permitted us to conduct biochemical and structural studies of starting materials and subsequent population doublings. Electron microscope evaluation of both initial monolayers and cell suspensions showed cultures to be composed of a single cell type. These cells had microvilli on their free or apical surface. Subsequent population doubling level 1 up to 5 exhibited the same structures. They contained lamellar inclusions, which are typical of Type II alveolar epithelial cells. Fetal lung (age 18 to 20 wk) cell suspensions processed for electron microscopy before culturing showed cells to be undifferentiated, epithelial-like with small microvilli along cell borders, and with desmosomes at cell junctions. Lamellar inclusions were not observed in these cells. Ultrastructural studies of the cultured epithelial cells demonstrated that the lamellar inclusions had a slightly positive reaction when tested for acid phosphatase. Phospholipid analysis of these lung epithelial cells showed a phospholipid composition consistent with that found in surfactant-containing Type II cells. Cultured epithelial cells stained with phosphine 3-R demonstrated a green fluorescent cytoplasm and nucleus with brightly fluorescent yellow-orange perinuclear particles. The preceding characterization of these cells leads us to conclude that they exhibit structural and biochemical features commensurate with Type II epithelial cells from human lung. Moreover, these selection techniques applied to the isolation of human lung Type II cells from the tissue permit us to study the differentiative function of these cells routinely under conditions of growth in vitro.

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
在无胶原微环境中生长的人II型肺泡细胞的生长特征、形态和磷脂组成
人肺上皮细胞已被分离并在纯培养中维持,并在培养过程中进行了表征。在培养的头48小时内,通过选择性胰蛋白酶化去除残留的成纤维细胞,原代培养中残留的上皮细胞生长到融合密度。当培养达到约80%的融合度时,2代或更大的上皮细胞被连续亚代。这个程序允许我们对起始材料和随后的种群加倍进行生化和结构研究。电子显微镜对初始单层和细胞悬浮液的评价表明,培养物由单一细胞类型组成。这些细胞的游离表面或顶端有微绒毛。随后的种群倍增1级至5级表现出相同的结构。它们含有典型的II型肺泡上皮细胞的片状内含物。胚胎肺(18 - 20周)细胞悬液在培养前的电镜观察显示,细胞未分化,呈上皮样,细胞边缘有小微绒毛,细胞连接处有桥粒。在这些细胞中未观察到片状包涵体。培养上皮细胞的超微结构研究表明,在酸性磷酸酶检测中,片层包涵体有轻微的阳性反应。这些肺上皮细胞的磷脂分析显示磷脂组成与含有表面活性剂的II型细胞一致。培养的上皮细胞用磷化氢3-R染色,胞质和细胞核呈绿色荧光,核周颗粒呈明亮的黄橙色荧光。上述对这些细胞的表征使我们得出结论,它们具有与人肺II型上皮细胞相称的结构和生化特征。此外,这些选择技术应用于从组织中分离人肺II型细胞,使我们能够在体外生长条件下研究这些细胞的常规分化功能。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
自引率
0.00%
发文量
0
期刊最新文献
Serial propagation of adult human prostatic epithelial cells with cholera toxin. Synergistic effect of TPA and T-cell mitogens in nonmammalian vertebrates. Glycogen synthesis in serum-free cultured hepatocytes in response to insulin and dexamethasone. Stimulation and inhibition of myoblast differentiation by hormones. Drosophila cells and ecdysterone: a model system for gene regulation.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1