解剖学报 ›› 2019, Vol. 50 ›› Issue (6): 735-740.doi: 10.16098/j.issn.0529-1356.2019.06.006

• 细胞和分子生物学 • 上一篇    下一篇

兔心包液CD44和波形蛋白阳性细胞具有多项分化潜能

孙莹 孙攀文 郭志坤*   

  1. 新乡医学院河南省医用组织再生重点实验室,河南 新乡 453003
  • 收稿日期:2018-12-21 修回日期:2019-03-18 出版日期:2019-12-06 发布日期:2019-12-06
  • 通讯作者: 郭志坤 E-mail:gzk@xxmu.edu.cn
  • 基金资助:
    国家自然科学基金

CD44 and vimentin positive cells in rabbit pericardial fluid having multiple differentiation potentials in vitro

SUN Ying SUN Pan-wen GUO Zhi-kun*   

  1. He’nan Key Laboratory of Medical Tissue Regeneration, Xinxiang Medical University, He’nan Xinxiang 453003, China
  • Received:2018-12-21 Revised:2019-03-18 Online:2019-12-06 Published:2019-12-06
  • Contact: GUO Zhi-kun* E-mail:gzk@xxmu.edu.cn

摘要:

目的 探讨兔心包液内的细胞成分及其分化潜能,为心包液细胞基础研究和临床应用提供形态学依据。 方法 成年新西兰大白兔 30 只, 无菌开胸,用输液针软管抽取心包液,离心培养。取不同培养代次细胞,倒置显微镜下行细胞形态观察。利用免疫荧光技术对心包液细胞进行免疫显色,观察CD44、CD45、波形蛋白(vimentin)的表面标记。取第3代细胞进行成骨、成脂诱导。利用 PCR技术检测CD44、CD45、vimentin相关分子表达情况及分析表达量的差异。结果 成年兔心包液中含有形态较为均匀、生长状态良好、增殖能力强的细胞群。免疫荧光结果显示,CD44、vimentin阳性表达, 不表达CD45。成脂和成骨诱导后,可向成骨和成脂细胞分化。 结论 兔心包液中含有多项分化潜能的细胞,它们可能对心肌修复具有积极意义。

关键词: CD44, CD45, 波形蛋白, 心包液, 免疫荧光, 新西兰白兔

Abstract:

Objective To investigate the cellular components and differentiation potential of cells in rabbit pericardial fluid, and to provide morphological basis for basic research and clinical application of pericardial cells. Methods Thirty adult New Zealand white rabbits, after aseptic thoracotomy, the pericardial fluid mixture was extracted, the fluid cells were centrifuged, isolated and cultured.The pericardial cellular morphology in the different generations was observed under the inverted microscope(The immunofluorescence staining method was used in the present study in order to analyze the pericardial cells phenotypes).Their immunological phenotypes were analyzed by using immunofluorescence staining and the CD44, vimentin, CD45 and the number of cells positively expressed in the third generation cells were observed.The expression of CD44 and vimentin related molecules was detected by PCR. Results There was the cellular population with uniform morphology in the adult rabbit pericardial fluid. The cells with immunofluorescence positive staining for CD44 and vimentin were found in the pericardial fluid of rabbit, in addition, these cells possessed the immunofluorescence negative staining for CD45.After induction, they can differentiate into osteoblasts and adipocytes. Conclusion Rabbit pericardial fluid contains cells with multiple differentiation potentials, which may be of positive significance for myocardial repair.

Key words: CD44, CD45, Vimentin, Pericardial fluid, Immunofluorescence, New Zealand white rabbit