军事医学科学院院刊
軍事醫學科學院院刊
군사의학과학원원간
BULLETIN OF THE ACADEMY OF MILITARY MEDICAL SCIENCES
2010年
1期
34-36
,共3页
孟革%赵建%吕新怀%丁日高
孟革%趙建%呂新懷%丁日高
맹혁%조건%려신부%정일고
肺泡Ⅱ型上皮细胞%分离%纯化%原代培养%鉴定
肺泡Ⅱ型上皮細胞%分離%純化%原代培養%鑒定
폐포Ⅱ형상피세포%분리%순화%원대배양%감정
alveolar type Ⅱ epithelial cell%isolation%purification%primary culture%identification
目的 建立大鼠肺泡Ⅱ型上皮细胞(alveolar type Ⅱ epithelial cell, AT-Ⅱ)分离、纯化、原代培养及鉴定方法.方法 取Wistar大鼠肺脏,采用胰蛋白酶消化法分离AT-Ⅱ,通过差速离心法、红细胞裂解法、贴壁选择法、免疫黏附选择法纯化AT-Ⅱ,倒置相差显微镜下观察细胞形态,鞣酸染色法进行细胞鉴定.结果 与结论 AT-Ⅱ呈单个或岛状生长,细胞呈圆形或椭圆形,细胞浆内有大量的反差明显的细胞小颗粒,细胞核明显.鞣酸染色法鉴定阳性.通过改进的酶消化法获得的AT-Ⅱ生长状态良好,纯度高,符合体外实验要求.
目的 建立大鼠肺泡Ⅱ型上皮細胞(alveolar type Ⅱ epithelial cell, AT-Ⅱ)分離、純化、原代培養及鑒定方法.方法 取Wistar大鼠肺髒,採用胰蛋白酶消化法分離AT-Ⅱ,通過差速離心法、紅細胞裂解法、貼壁選擇法、免疫黏附選擇法純化AT-Ⅱ,倒置相差顯微鏡下觀察細胞形態,鞣痠染色法進行細胞鑒定.結果 與結論 AT-Ⅱ呈單箇或島狀生長,細胞呈圓形或橢圓形,細胞漿內有大量的反差明顯的細胞小顆粒,細胞覈明顯.鞣痠染色法鑒定暘性.通過改進的酶消化法穫得的AT-Ⅱ生長狀態良好,純度高,符閤體外實驗要求.
목적 건립대서폐포Ⅱ형상피세포(alveolar type Ⅱ epithelial cell, AT-Ⅱ)분리、순화、원대배양급감정방법.방법 취Wistar대서폐장,채용이단백매소화법분리AT-Ⅱ,통과차속리심법、홍세포렬해법、첩벽선택법、면역점부선택법순화AT-Ⅱ,도치상차현미경하관찰세포형태,유산염색법진행세포감정.결과 여결론 AT-Ⅱ정단개혹도상생장,세포정원형혹타원형,세포장내유대량적반차명현적세포소과립,세포핵명현.유산염색법감정양성.통과개진적매소화법획득적AT-Ⅱ생장상태량호,순도고,부합체외실험요구.
Objective To establish a method of isolation, purification, primary culture and identification of alveolar type Ⅱ epithelial cells(AT-Ⅱ).Methods The AT-Ⅱs were isolated from Wistar rats by trypsin,purified by differential centrifugation, erythrocyte spallation, differential adherence and immune adherence, and identified by observing the morphology of cultured cells under the inverted phase and tannic acid staining. Results and Conclusion The cultured primary AT-Ⅱs in vitro presented single or island form growth, and their shapes were round or elliptical. A great deal of fine particles showed sharp contrast, and were observed in intracytoplasm. The cell nuclei were clear. They were positive for tannic acid staining.The primary culture AT-Ⅱs obtained from improved isolation and purification have good growth state and purity, and are suitable for research in vitro.
