A novel method for endothelial cell isolation

  • Authors:
    • Qiqi Mao
    • Xianing Huang
    • Jian He
    • Wei Liang
    • Yi Peng
    • Jing Su
    • Yingying Huang
    • Zixi Hu
    • Xiaoling Lu
    • Yongxiang Zhao
  • View Affiliations

  • Published online on: December 16, 2015     https://doi.org/10.3892/or.2015.4490
  • Pages: 1652-1656
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

The present study aimed to develop a quick and efficient method for purification of newborn endothelial cells from tumor tissues. Fresh tissues were separated from C57BL/6 mice bearing tumors derived from mouse lung cancer Lewis cells, fully minced and divided into two parts. One part was subjected to collagenase type I digestion with a vortex to form a single-cell suspension, while another part was digested but without a vortex. Then, the CD105+ cells were isolated using anti-CD105 antibody-coated Dynabeads. The isolated CD105+ cells were grown in culture medium and examined for the surface expression of CD105 by a fluorescence-activated cell sorter (FACS). The uptake of acetylated LDL and the ability to maintain capillary tube-like structure formation in the CD105+ cells were also examined by Dil-Ac-LDL uptake assay and tube formation assay. The expression of tumor newborn endothelial cells (CD105+) was tested in Lewis xenografts by immunohistochemistry. The number of cells which were obtained by the digestion process with a vortex was 5.70±0.23x104 much higher than the number without a vortex (0.32±0.04x104) (P<0.01). The purity of CD105+ cell digestion with a vortex was significantly higher than that without a vortex. Dil-Ac-LDL uptake assay and tube formation assay confirmed that the CD105+ cells digested with a vortex exhibited typical functions of endothelial cells. In conclusion, the CD105+ cells isolated by the new method had high purity and displayed features of vascular endothelial cells. The modified method provides CD105+ cells with superior conditions for mechanistic research on the development of vessel-based disease.
View Figures
View References

Related Articles

Journal Cover

March-2016
Volume 35 Issue 3

Print ISSN: 1021-335X
Online ISSN:1791-2431

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Mao Q, Huang X, He J, Liang W, Peng Y, Su J, Huang Y, Hu Z, Lu X, Zhao Y, Zhao Y, et al: A novel method for endothelial cell isolation. Oncol Rep 35: 1652-1656, 2016
APA
Mao, Q., Huang, X., He, J., Liang, W., Peng, Y., Su, J. ... Zhao, Y. (2016). A novel method for endothelial cell isolation. Oncology Reports, 35, 1652-1656. https://doi.org/10.3892/or.2015.4490
MLA
Mao, Q., Huang, X., He, J., Liang, W., Peng, Y., Su, J., Huang, Y., Hu, Z., Lu, X., Zhao, Y."A novel method for endothelial cell isolation". Oncology Reports 35.3 (2016): 1652-1656.
Chicago
Mao, Q., Huang, X., He, J., Liang, W., Peng, Y., Su, J., Huang, Y., Hu, Z., Lu, X., Zhao, Y."A novel method for endothelial cell isolation". Oncology Reports 35, no. 3 (2016): 1652-1656. https://doi.org/10.3892/or.2015.4490