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TISSUE-SPECIFIC STEM CELLS |
aCenter for Gene Therapy, Tulane University Health Sciences Center, New Orleans, Louisiana, USA;
bDepartment of Medicine, Cardiovascular Research Institute, University of Vermont, Colchester, Vermont, USA
Key Words. Adult bone marrow stem cells • Stem cell culture • Mesenchymal stem cells • Immune system • Experimental models • Ex vivo expansion • Engraftment
Correspondence: Carl A. Gregory, Ph.D., Center for Gene Therapy, Tulane University Health Sciences Center, 1430 Tulane Avenue, New Orleans, Louisiana 70112, USA. Telephone: 504-988-7176; Fax: 504-988-7710; e-mail: cgregory{at}tulane.edu
Received December 5, 2005;
accepted for publication May 30, 2006.
First published online in STEM CELLS EXPRESS June 8, 2006.
Human mesenchymal stem cells (hMSCs), also referred to as multipotent stromal cells, are currently being applied in clinical trials for bone diseases, graft versus host disease, and myocardial infarction. However, the standard growth medium for hMSCs contains 10%–20% fetal calf serum (FCS), and FCS is strongly immunogenic in both rodents and humans. Previously, we reported that by a sensitive fluorescence-based assay, 7–30 mg of internalized FCS is associated with 108 hMSCs, a dosage that will probably be needed for most therapies. We also found that a brief culture in medium containing autologous 20% adult human serum (AHS) or autologous 10% AHS supplemented with growth factors (AHS+) reduced the contamination by more than 99.9%. We have now extensively characterized the culture conditions and shown that hMSC expansion is possible using heterologous 20% AHS or heterologous 10% AHS+. The uptake of FCS is an active process that acts to concentrate contamination in the cells even under low serum conditions (2% FCS) but can be actively displaced by incubation of the cells in medium with AHS. Rat MSCs (rMSCs) can be expanded under similar conditions using supplemented heterologous adult rat serum (ARS+). After expansion in FCS, a further 8 days of culture with ARS+ significantly improves the viability of the rMSCs in vivo after encapsulation in fibrin followed by subcutaneous implantation in rats. Our results have the potential to dramatically improve cellular and genetic therapies using hMSCs.
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