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Trophic factor induction of human umbilical cord blood cells in vitro and in vivo

Ning Chen1, Siddharth Kamath2, Jennifer Newcomb1, Jennifer Hudson1, Svitlana Garbuzova-Davis3, Paula Bickford4, Cyndy Davis-Sanberg5, Paul Sanberg6, Tanja Zigova1 and Alison Willing7

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The mononuclear fraction of human umbilical cord blood (HUCBmnf) is a mixed cell population that multiple research groups have shown contains cells that can express neural proteins. In these studies, we have examined the ability of the HUCBmnf to express neural antigens after in vitro exposure to defined media supplemented with a cocktail of growth and neurotrophic factors. It is our hypothesis that by treating the HUCBmnf with these developmentally-relevant factors, we can expand the population, enhance the expression of neural antigens and increase cell survival upon transplantation. Prior to growth factor treatment in culture, expression of stem cell antigens is greater in the non-adherent HUCBmnf cells compared to the adherent cells (p < 0.05). Furthermore, treatment of the non-adherent cells with growth factors, increases BrdU incorporation, especially after 14 days in vitro (DIV). In HUCBmnf-embryonic mouse striata co-culture, a small number of growth factor treated HUCBmnf cells were able to integrate into the growing neural network and express immature (nestin and TuJ1) and mature (GFAP and MAP2) neural markers. Treated HUCBmnf cells implanted in the subventricular zone predominantly expressed GFAP although some grafted HUCBmnf cells were MAP2 positive. While short-term treatment of HUCBmnf cells with growth and neurotrophic factors enhanced proliferative capacity in vitro and survival of the cells in vivo, the treatment regimen employed was not enough to ensure long-term survival of HUCBmnf-derived neurons necessary for cell replacement therapies for neurodegenerative diseases.


PACS

87.85.Lf Tissue engineering

87.17.-d Cell processes

87.17.Ee Growth and division

87.19.L- Neuroscience

Subjects

Medical physics

Biological physics

Dates

Issue 2 (June 2007)

Received 3 August 2006, accepted for publication 20 February 2007

Published 4 April 2007



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