Limiting-dilution transplantation assays in mammary stem cell studies Journal Article
| Authors: | Illa-Bochaca, I.; Fernandez-Gonzalez, R.; Shelton, D. N.; Welm, B. E.; Ortiz-de-Solorzano, C.; Barcellos-Hoff, M. H. |
| Article Title: | Limiting-dilution transplantation assays in mammary stem cell studies |
| 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. |
| Keywords: | methodology; cell proliferation; mouse; animal; cytology; animals; mice; transplantation; stem cell; staining; epithelium cell; epithelial cells; stem cells; centrifugation; udder; mammary glands, animal; cell separation; cell transplantation; adipose tissue; staining and labeling; article |
| Journal Title: | Methods in Molecular Biology |
| Volume: | 621 |
| ISSN: | 1064-3745 |
| Publisher: | Humana Press Inc |
| Date Published: | 2010-01-01 |
| Start Page: | 29 |
| End Page: | 47 |
| Language: | English |
| DOI: | 10.1007/978-1-60761-063-2_2 |
| PUBMED: | 20405357 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Book Chapter 2 in "Protocols for Adult Stem Cells" (ISBN: 978-1-60761-062-5) -- Export Date: 6 March 2020 -- Source: Scopus |
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