Isolation and primary culture of typeⅡ alveolar epithelial cells from fetal rat lung

FU Hong-min; et al

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Isolation and primary culture of typeⅡ alveolar epithelial cells from fetal rat lung

更新时间：2024-01-17

- Isolation and primary culture of typeⅡ alveolar epithelial cells from fetal rat lung
- Journal of Chongqing Medical University Vol. 34, Issue 5, Pages: 586-589(2009)
- 作者机构：
  
  重庆医科大学附属儿童医院儿科重症监护室
- 作者简介：
- 基金信息：
- DOI：
  CLC： R329
- Published：2009，
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[1]付红敏,许峰,黄波,杨鸣,符跃强,方芳,匡凤梧.胎鼠肺泡Ⅱ型上皮细胞的分离及原代培养[J].重庆医科大学学报,2009,34(05):586-589.

FU Hong-min, et al. Isolation and primary culture of typeⅡ alveolar epithelial cells from fetal rat lung. [J]. Journal of Chongqing Medical University 34(5):586-589(2009)
[1]付红敏,许峰,黄波,杨鸣,符跃强,方芳,匡凤梧.胎鼠肺泡Ⅱ型上皮细胞的分离及原代培养[J].重庆医科大学学报,2009,34(05):586-589. DOI：

FU Hong-min, et al. Isolation and primary culture of typeⅡ alveolar epithelial cells from fetal rat lung. [J]. Journal of Chongqing Medical University 34(5):586-589(2009) DOI：

摘要

目的:探讨胎鼠肺泡Ⅱ型上皮细胞（Alveolar epithelial cell typeⅡ

AECⅡ）的分离、纯化及原代培养方法

为有关胎儿肺发育及新生儿肺部疾病的研究奠定基础。方法:采用胰酶结合胶原酶的消化方法

分离肺组织细胞成份

然后经差速离心和差速贴壁的方法纯化AECⅡ

进行原代培养;通过台盼蓝染色检测细胞活力

倒置相差显微镜观察细胞生长特点及形态特征

透射电镜鉴定

改良巴氏染色检测细胞纯度以及免疫荧光技术检测表面蛋白C（Surfactant proteinc

SPC）的表达。结果:每3～5只胎鼠可获得AECⅡ（36±5）×106

活力98%±2%。镜下观察原代AECⅡ呈岛状生长

外观呈多边形或立方形。透射电镜可见特征性的板层小体

改良巴氏染色见胞质内有较多颗粒

纯度为96%±3%

呈现SPC绿色免疫荧光的细胞占96%以上。结论:利用胰酶和胶原酶消化

以及差速离心和差速贴壁的方法可成功分离出高产量、高纯度的胎鼠AECⅡ

能满足体外进一步实验的需要。

Abstract

Objective:Todevelop the isolation and purification technology for typeⅡalveolar epithelial cells(AECⅡ)of fetal rat

in order to provide experimental means for the study of lung development or neonatal lung disease.Methods:Lung tissues of 19-day fetal rats were digested with trypsin and collagenase

then purified for AECⅡwith different centrifugal force and repeated attachment.Cell viability was evaluated by typran inclusion dying before plated into six-well flask.Growth status and shape of attached cells were observed with inverted phase contrast microscope.AECⅡwere identified by electron microscopy and its percentage was assessed by modified Papanicolaou dying and immunofluorescence

the latter aiming to detect expression of surfactant protein C(SPC)in AECⅡ.Results:The total amount of cells we harvested from 3 to5 fetal rat was(36±5)×106 with a viability of 98%±2%.Cells were like polygonal and connected like island under microscope.AECⅡwas ascertained by lamellar bodies found in cytoplasma under electron microscope and its percentage was 96%±3%identified by modified Papanicolaou dying

the same as the result by immunofluorescence for SPC.Conclusion:Highly Purified and viable AECⅡcould be achieved by our methods and the cell model could be used in further study.

关键词

胎鼠肺泡Ⅱ型上皮细胞原代培养分离

Keywords

Fetal ratAlveolar epithelial cell typeⅡprimary cultureIsolation

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