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

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
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

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