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Ex Vivo Expansion of Stem Cells from Umbilical Cord Blood: Expression of Cell Adhesion Molecules

Chemotherapy and Stem Cell Biology Division, Cancer Research Institute, Tata Memorial Centre, Parel, Mumbai, India

Key Words. Cord blood • CD34⁺ cells • Expansion • Cell adhesion molecules • CAMs

S.G.A. Rao, Ph.D., Reliance Life Sciences, Sir H.N. Hospital, PG Chowk, RajaRam Mohan Roy Road, Mumbai – 400 004, India. Telephone: 91-22-3810707 ext. 503; Fax: 91-22-3889390; e-mail: sgarao{at}vsnl.net

Expansion of stem cells from cord blood has been demonstrated to increase the numbers of CD34⁺ cells, CD34⁺ subsets, long-term culture-initiating cells, and severe combined immunodeficient mouse, repopulating cells. However, reports suggest that the ex vivo expanded population behaves differently than freshly isolated cells and shows a delayed or diminished engraftment. In this study, we investigated the effects of the cytokines flt3 ligand, stem cell factor, and thrombopoietin on expansion of CD34⁺ and CD34⁺/CD38^- cells. In addition, we studied the expression of adhesion molecules, very late activation antigen-4 (VLA-4) and leukocyte function antigen-1 (LFA-1), on CD34⁺ cells from cord blood by flow cytometry. We also looked at the expression of an adhesion receptor, namely, vascular cell adhesion molecule-1 (VCAM-1) on bone marrow stromal cells by Western blot analysis after exposure to low dose irradiation. After culturing for 7 days, increases in the absolute numbers of CD34⁺, CD34⁺/CD38^-, CD34⁺/VLA-4⁺, and CD34⁺/LFA-1⁺ cells were 5.67 ± 2.91 (mean ± standard deviation) fold, 7.21 ± 4.38 fold, 99.56 ± 101.5 fold, and 101.39 ± 83.25 fold, respectively. There was a transient upregulation in the expression levels of VCAM-1 on stromal cells, which peaked at 4 hours. Though there was an increase in the absolute numbers of CD34⁺ cells expressing the adhesion molecules, the expression levels (antigen density) of the adhesion molecules on the CD34⁺ cells remained unaffected.

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				P. Denning-Kendall, S. Singha, B. Bradley, and J. Hows Cytokine Expansion Culture of Cord Blood CD34+ Cells Induces Marked and Sustained Changes in Adhesion Receptor and CXCR4 Expressions Stem Cells, January 1, 2003; 21(1): 61 - 70. [Abstract] [Full Text] [PDF]