Isolation and primary culture of human proximal tubule cells

Vesey, David A., Qi, Weier, Chen, Xingming, Pollock, Carol A. and Johnson, David W. (2009). Isolation and primary culture of human proximal tubule cells. In John M. Walker, T. D. Hewitson and G. J. Becker (Ed.), Kidney Research: Experimental Protocols (pp. 19-24) Clifton, NJ, U.S.A.: Humana Press. doi:10.1007/978-1-59745-352-3_2

Author Vesey, David A.
Qi, Weier
Chen, Xingming
Pollock, Carol A.
Johnson, David W.
Title of chapter Isolation and primary culture of human proximal tubule cells
Title of book Kidney Research: Experimental Protocols
Place of Publication Clifton, NJ, U.S.A.
Publisher Humana Press
Publication Year 2009
Sub-type Research book chapter (original research)
DOI 10.1007/978-1-59745-352-3_2
Series Methods in Molecular Biology
ISBN 9781588299451
Editor John M. Walker
T. D. Hewitson
G. J. Becker
Volume number 466
Chapter number 2
Start page 19
End page 24
Total pages 6
Total chapters 18
Collection year 2010
Language eng
Subjects 0601 Biochemistry and Cell Biology
060103 Cell Development, Proliferation and Death
Formatted Abstract/Summary
Primary cultures of renal proximal tubule cells (PTC) have been widely used to investigate tubule cell function. They provide a model system where confounding influences of renal haemodynamics, cell heterogeneity, and neural activity are eliminated. Additionally they are likely to more closely resemble PTC in vivo than established kidney cell lines, which are often virally immortalised and are of uncertain origin. This chapter describes a method used in our laboratories to isolate and culture pure populations of human PTC. The cortex is dissected away from the medulla and minced finely. Following collagenase digestion, the cells are passed through a sieve and separated on a Percoll density gradient. An almost pure population of tubule fragments form a band at the base of the gradient. Cultured in a hormonally defined serum-free growth media, they form a tightly packed monolayer that retains the differentiated characteristics of PTC for up to three passages.
© Humana Press, Totowa, NJ
Keyword Renal proximal tubule cells
Hormonally defined serum free medium
Percoll gradient
Polarised monolayers
Q-Index Code B1
Q-Index Status Confirmed Code
Additional Notes Published in Part 1: "Isolation and Propagation of Cell Populations". ISSN: 1064-3745 (Print) 940-6029 (Online).

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Created: Wed, 07 Apr 2010, 13:28:22 EST by Fiona Mactaggart on behalf of Medicine - Princess Alexandra Hospital