The basic research for the establishment of the donor for the neural transplantation with the miniature swine neuroepitherial stem cells.

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The tissue and cell dissociation culture of the embryonic day 10.5 (E10.5) Wister rats mesencephalic neural plate, consisted wife neuroepitheral stem cells (NESCs). were cultured in vitro under various conditions and were analyzed 7 days after the primary culture. When they were cultured as a tissue, almost all cells differentiated into neurofilament (NF) positive neuron extending long neurites in a serum-free and serum containing medium. But, there were no astrocyte expressed glial fibrially acidic protein (GFAP). For the evaluation of the naerochemical marker, many glutamate positive cells, few γ-aminoburyric acid (GABA) and tyrosine bydroxylase (TH) positive cells were there. On the other hand, when they were plated in a dissociation culture, neurosphere generated 7 days after the primary cell culture in the medium containing fibroblast growth factor-2 (FGF-2). The FGF-2 responsive neurosphere were positively stained by nestin and able to maintain. Under the condition of differentia … More tion all cells of the primary neurosphere differentiated into NF positive neuron, not into GFAP positive cell, in the differentiated neurosphere, many glutamate positive cells few GABA and TH positive cells were there as same as the tissue culture. These indicated that the rat mesencephalic neural plate cells can keep the neuronal differentiation tendency until one week by FGF-2 administration.We also analyzed immunohisochemically the biological nature of the mesensephalic neural plate and FGF-2 responsive neurospheres transplanted into the normal adult rat brain. The viable graft in the host brain was observed for several weeks after transplantation and many Hu-positive neuronal cells and a few GFAP-positive cells were seen in the graft Especially in the FGF-2 responsive graft marry. Hu-positive neuronal cells and a few nestin-positive cells were observed, but no GFAP-positive cells could be identified 2 weeks after transplantation. These results suggested that NESCs demonstrated a propensity to differentiate into neurons in the adult brain environment.The mesencephalic neural tube tissue graft from E17-18 swine embryos implanted immediately into the strianum of the Parkinsonian model rat were observed for several weeks with 1-week administration of the immunosuppressants after transplantation only. The swine derived grafts had many NF-positive and TH-positive cells in the host rat striatum. An electron microscopic study revealed both efferent and afferent synaptic formations in the donor-derived immature neurons or TH-positive donor cells in the grafts. Myelimated axons, bath positive and negative for swine-specific NF antibody, were mingled together in the graft.Both mini-swine derived NESCs and rat derived NESCs survived in Parkinsonian rat striatum at 8 weeks after transplantation. The rat NESCs grafted rat showed complete recovery from Parkinsonian behavioral defects in amphetamine-induced rotation test. On the other hand, the mean number of amphetamine-induced rotations made by the mini-swine NESCs grafted rats expressed as a percentage of the number before grafting had not changed until 4 weeks after grafting but decreased to 30% at 8 weeks after grafted in the grafts of both mini-swine NESCs and rat NESCs, many TH positive donor-derived cells were there 2 weeks after transplantation for the immunohistolgical evaluation.NESCs are therefore an attractive candidate as a source of donor material for intracerebral grafting in neural circuitry reconstruction Less