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Limiting-Dilution Transplantation Assays in Mammary Stem Cell Studies

Authors :
Carlos Ortiz-de-Solorzano
Bryan E. Welm
Mary Helen Barcellos-Hoff
Rodrigo Fernandez-Gonzalez
Dawne N. Shelton
Irineu Illa-Bochaca
Source :
Methods in Molecular Biology ISBN: 9781607610625
Publication Year :
2010
Publisher :
Humana Press, 2010.

Abstract

Mammary reconstitution assays can be used to measure the stem cell frequency within an epithelial population by transplanting increasingly diluted single-cell preparations of the population of interest. There are fundamental steps in the single-cell isolation protocol which are directly related to the number of single epithelial cells obtained. Once single-cell suspensions have been obtained, serial dilutions are prepared and transplanted into the cleared fat pads of the host mice. After 8-10 weeks, the transplanted fat pads are reevaluated for the presence of epithelial outgrowths. Based on the frequency of no outgrowth for each one of the transplanted dilutions, it is possible to estimate the frequency of mammary repopulating cells present in a given cell population. Here, we give details on how to carry out all these steps.

Details

ISBN :
978-1-60761-062-5
ISBNs :
9781607610625
Database :
OpenAIRE
Journal :
Methods in Molecular Biology ISBN: 9781607610625
Accession number :
edsair.doi...........909a045fdef9b4c1d2b0269758ee5b3a