Isolation and epithelial co-culture of mouse renal peritubular endothelial cells

BMC Cell Biology

Guoping Zheng²  David C Harris²  Steve I Alexander³  Yuan Min Wang³  Yiping Wang²  Ya Wang²  Qi Cao²  Tania Tsatralis²  Yun Zhang⁴  Hong Zhao¹  Ye Zhao² 

[1] Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, PR China;Centre for Transplant and Renal Research, Westmead Millennium Institute, The University of Sydney, Sydney, NSW, Australia;Centre for Kidney Research, Children’s Hospital at Westmead, Sydney, NSW, Australia;Experimental Centre of Science and Research, The First Clinical Hospital of Shanxi Medical University, Taiyuan, PR China

关键词: Vascular endothelial growth factor;    Co-culture;    CD146;    Tubular epithelial cells;    Peritubular endothelial cells;   

Others  :  1088716
DOI  :  10.1186/s12860-014-0040-6

 received in 2014-03-04, accepted in 2014-10-16,  发布年份 2014

【 摘 要 】

Background

Endothelial-mesenchymal transition (EndoMT) has been shown to be a major source of myofibroblasts, contributing to kidney fibrosis. However, in vitro study of endothelial cells often relies on culture of isolated primary endothelial cells due to the unavailability of endothelial cell lines. Our recent study suggested that peritubular endothelial cells could contribute to kidney fibrosis through EndoMT. Therefore, successful isolation and culture of mouse peritubular endothelial cells could provide a new platform for studying kidney fibrosis. This study describes an immunomagnetic separation method for the isolation of mouse renal peritubular endothelial cells using anti-CD146 MicroBeads, followed by co-culture with mouse renal proximal tubular epithelial cells to maintain endothelial phenotype.

Results

Flow cytometry showed that after isolation and two days of culture, about 95% of cells were positive for endothelial-specific marker CD146. The percentage of other cells, including dendritic cells (CD11c) and macrophages (F4/80), was less than 1%. Maintenance of endothelial cell phenotype required vascular endothelial growth factor (VEGF) and co-culture with mouse proximal tubular epithelial cells.

Conclusion

In this study, we established a method for the isolation of mouse renal peritubular endothelial cells by using immunomagnetic separation with anti-CD146 MicroBeads, followed by co-culture with mouse renal proximal tubular epithelial cells to maintain phenotype.

【 授权许可】

   
2014 Zhao et al.; licensee BioMed Central Ltd.

【 图 表 】

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