Stem cell culture medium and method of using said medium and the cells

Patent metadata

Field	Value
Publication number	US-9523080-B2
Application number	US-201514591819-A
Country	US
Kind code	B2
Filing date	Jan 7, 2015
Priority date	Nov 4, 2003
Publication date	Dec 20, 2016
Grant date	Dec 20, 2016

Abstract

The present invention relates to methods and compositions concerning isolation of proliferating cells. In particular, the invention regards enrichment of stem cells in a mixture of stem cells and non-stem cells, wherein the non-stem cells may be differentiated cells. The invention exploits 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.

First claim

What is claimed is:

1. A method of isolating CD34+ stem cells from human peripheral blood, comprising the steps of:
culturing cells obtained from human peripheral blood 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.

2. The method of claim 1 , wherein the cells were obtained from human peripheral blood by flow cytometry.

3. The method of claim 1 , wherein the first liquid cell culture medium lacks matrix and feeder cells.

4. The method of claim 1 , wherein the subsequent liquid cell culture medium lacks matrix and feeder cells.

5. The method of claim 1 , wherein the passaging occurs about once a week.

6. The method of claim 1 wherein the passaging occurs more than once a week.

7. The method of claim 1 , wherein the isolated cells express Oct4, Neurofilament H, Sca-1, AA4.1, cKit, CD45, HMBP, Pax-6, HuC/HuD, NeuN, GAD65, CNPase, MOSP, MBP2, galactocerebroside, NG2, or a combination thereof.

8. The method of claim 1 , further comprising the step of delivering one or more of the stem cells to an individual.

Assignees

• Univ Maryland
• The United States As Represented By The Dept Of Veterans Affairs
• Us Veterans Affairs

Inventors

• Trisler G David
• Bever Jr Christopher T
• Goolsby James E
• Pessac Bernard M

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• A61P3/10

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• A61P35/02

  specific for leukemia · CPC title

• A61P9/00

  Drugs for disorders of the cardiovascular system · CPC title

• A61P25/16

  Anti-Parkinson drugs · CPC title