Isolation and characterization of hematopoietic stem cells from human umbilical cord blood for cell therapy

Durga Prameela G., T.N.V. K.V. Prasad*, M. Nagalakshmi Devamma, E. K. Elumalai, P. M. Ayyasamy, T. Kiran Reddy


Alternatives to organ transplantation for treatment of certain liver diseases are currently under investigation. Whole liver, split liver and related living donor liver transplantation are clinically safe and well established procedures for the treatment of end-stage liver failure. However, organ donor shortage and the need of lifelong immunosuppressive treatment are still the major limitations of all these options. Therefore, the development of cell based therapeutic strategies for liver disease is under experimental evaluation. Umbilical cord blood was collected and isolated the mononucleated cell by the help of ficoll hypaque centrifugation. The collected samples were purified for CD 34+ hematopoietic stem cells using MACS. These purified hematopoietic stem cells were counted and morphologically characterized by FACS. Freshly isolated CD 34+ cells were cultured in hepatocyte differentiation media; contain growth factor of HGF and FGF-4. These growth factors induced the differentiation of hematopoietic stem cells into hepatocyte like cells. The cell density played a significant role in effecting hepatic differentiation. Optimal cell density was found to be 1.0 to 1.3X10⁴ cells /cm². After 14 inductions, cuboidal morphology, which is characteristic of hepatocytes were observed and cells also expressed by marker genes specific for liver cells in a time depended manner.


Umbilical Cord Blood; Hematopoietic Stem Cells; Hepatocyte; Marker Genes.

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Sherlock S, Lesh R, Reutter W, Experimental hepatitis induced by D-galactosamine. Exp. Mol. Pathol. 1994; 9: 279-90.

Starzl J, Schoemaker L, Gauthier T, Cerebral metabolic and histological effects of thiocetamide-induced liver failure, Am J. Physiol, 1990; 265: G572-8.

Grande JW, Haugvicova R, Albgrecht J, Decreased potassium stimulated release of [³H] D aspartate from hippocampal slices distinguishes encephalopathy related to acute liver failure from that induced by simple hyperammonemia, Brain Res., 1992, 567, 165-8.

Rubistein P, Crrier C, Scaradavou A, Kurtzberg J, Adamson J, Migliaccio AR, Berkovitz LR, Cabbad M, Dobrila LN, Taylor PE, Rosenfield RE and Stevens CE, 1998; Outcomes among 562 recipients of placental-blood transplants from unrelated donors. N Engl J Med; 339: 1565-1577.

Wagner JE, Umbilical cord blood transplantation. Transfusion; 1995; 35: 619-621.

Heimfeld S, HLA-identical stem cell transplantation: is there an optimal CD34 cell dose? Bone Marrow Transplant; 2003, 31: 839-846.

Appelbaum FR, Marrow transplantation for hematologic malignanices, a brief review of current status and future prospects, Semin Hematol; 1988, 25 (suppl 3) 16-22.

Gordon MY, Greaves MF, Physiological mechanisms of stem cell regulation in bone marrow transplantation and haemopoiesis, Bone Marrow Transplant; 1989; 4: 335-8.

McAdams TA, Miller WM and Papoutsakis ET, Hematopoietic cell culture therapies: cell culture consideration. Trends Biotechnol; 1996; 14: 388-396.

Goodwin HS, Levenson SM, Novikooff PM, Survival, organization and function of micro carrier attached hepatocytes transplanted in rats. Proc Natl Acad Sci USA, 2001; 83: 7475-7479.

Bornstein R, Flores AI, Montalban MA, DEL Rey MJ, Del Rey MJ, De la Serna J, Gilsanz FA, modified cord blood collection method achieves sufficient cell levels for transplantation in most adult patients, Stem cells; 2005; 23: 324-334.

Yang YE, Ha CW, Jung M, Jin HJ, Lee M, Song H, Choi S, Oh W, Yang YS, Mesenchymal stem/progenitor cells developed in cultures from umbilical cord blood. Cytotherapy, 2004; 6: 476-486.

HC Figel, Claudius Lang, U Kneser, W Lambrecht, A R, Zander, D Kluth, Fetal and adult liver stem cells for liver regeneration and tissue engineering. Stem cells; 2000, 55: 71-74.

Ruhnke M, Ungefroren H, Zehle G, Bader M, Kremer B, Fandrich F, Long-term culture and differentiation of rat embryonic stem cell-like cells into neuronal, glial, endothelial, and hepatic lineages. Stem cells; 2003; 21: 428-436.



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