移植
干细胞
限制
生物
细胞生物学
医学
内科学
工程类
机械工程
作者
Irineu Illa-Bochaca,Rodrigo Fernández-González,Dawne N. Shelton,Bryan E. Welm,Carlos Ortíz-de-Solórzano,Mary Helen Barcellos‐Hoff
出处
期刊:Methods in molecular biology
日期:2010-01-01
卷期号:: 29-47
被引量:18
标识
DOI:10.1007/978-1-60761-063-2_2
摘要
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.
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