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Xenotransplantation of embryonic stem cell-derived motor neurons into the developing chick spinal cord.

Methods in molecular biology (Clifton, N.J.) (2008-12-18)
Hynek Wichterle, Mirza Peljto, Stephane Nedelec
RESUMEN

A growing number of specific cell types have been successfully derived from embryonic stem cells (ES cells), including a variety of neural cells. In vitro generated cells need to be extensively characterized to establish functional equivalency with their in vivo counterparts. The ultimate test for the ability of ES cell-derived neurons to functionally integrate into neural networks is transplantation into the developing central nervous system, a challenging technique limited by the poor accessibility of mammalian embryos. Here we describe xenotransplantation of mouse embryonic stem cell-derived motor neurons into the developing chick neural tube as an alternative for testing the ability of in vitro generated neurons to survive, integrate, extend axons, and form appropriate synaptic contacts with functionally relevant targets in vivo. Similar methods can be adapted to study functionality of other mammalian cells, including derivatives of human ES cells.

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Sigma-Aldrich
Proteína LIF recombinante de ratón ESGRO®, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^7 units/ml.
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Purmorphamine, A cell-permeable activator of Hedgehog signaling that induces osteoblast differentiation of multipotent mesenchymal progenitor cells C3H10T1/2 (EC₅₀ = 1 µM).