GENERATION OF DOPAMINE NEURONS FROM HUMAN EMBRYONIC STEM CELLS IN VITRO
I. V. Kozhucharova,1 I. I. Fridlyanskaya, V. I. Zemelko, Z. V. Kovaleva,
N. A. Pugovkina, L. L. Alekseenko, M. V. Charchenko, A. N. Shatrova,
N. D. Aksenov, T. M. Grinchuk, S. V. Anisimov, N. N. Nikolsky
Institute of Cytology RAS, St. Petersburg;
1 e-mail: kojuxarova@mail.ru
The aim of the study was to generate dopaminergic (DA) neurons from human embryonic stem cells (ESC)
in vitro. It was shown that human ESCs are able to differentiated into DA neurons without co-culture with stromal
cells. Terminal differentiation into DA neurons was reached by successive application of noggin and bFGF
growth factors on collagen and matrigel substrates during 3—4 weeks. Differentiation efficiency was evaluated
by the number of colonies with cells expressing tyrosine hydroxylase (TH), a DA neuron marker, and by the
number of TH-positive cells in cell suspension using flow cytometry. No cells with pluripotent markers were detected
in DA-differentiated cultures. It makes possible to propose that the protocol of human ESC differentiation
might be applied to generate DA neurons for their transplantation into the animals modeling neurodegenative
(Parkinson) disease without the risk of tumor growth.
Key words: human embryonic stem cells, neuronal differentiation, dopaminergic neurons, immunocytochemistry
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