Stem cell culture medium and method of using said medium and the cells
AssigneesUniversity of Maryland, Baltimore, The United States of America as Represented by the Dept. of Veterans Affairs
InventorsG. David Trisler, Christopher T. Bever, Jr., James E. Goolsby, Bernard M. Pessac
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Abstract
Methods and compositions are provided for isolation of proliferating cells. In particular, the methods enrich stem cells in a mixture of stem cells and non-stem cells, and in some cases the non-stem cells may be differentiated cells. The methods exploit the non-adherent property of stem cells, as opposed to the adherent property of differentiating cells, by serially passaging the suspended cells in liquid media.
Claims8
A method of isolating CD34+ stem cells from a plurality of bone marrow cells, comprising the steps of:
- culturing bone marrow cells in a first liquid cell culture medium that lacks serum, but that comprises interleukin-3, interleukin-6, stem cell factor, and beta-mercaptoethanol;
- successively passaging cells in suspension from the first medium to a subsequent liquid cell culture medium that lacks serum, but that comprises interleukin-3, interleukin-6, stem cell factor, and beta-mercaptoethanol, wherein the passaging occurs about once a week or more than once a week and the successive passaging steps occur over the course of three to four weeks;
- and
- isolating the cells in suspension from the final medium.
Dependencies are inferred from the claim text.