1987 Fiscal Year Final Research Report Summary
Replantation of caltured Schwann cells
| Project/Area Number |
61480332
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
麻酔学
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| Research Institution | Nagoya University School of Medicine |
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Principal Investigator |
MIYAMOTO Yoshihiro Department of Orthopaedic durgery, Hiroshima University School of Medicine, 医学部, 教授 (50028669)
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| Co-Investigator(Kenkyū-buntansha) |
貝沼 関志 名古屋大学, 医学部, 講師 (90135335)
石川 直久 名古屋大学, 医学部, 助教授 (80109321)
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| Project Period (FY) |
1986 – 1988
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| Keywords | Schwann cells / Dorsal root ganglia / Neurite outgrowth / Fibroblasts / Sciatic nerves / 星状神経節 |
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| Research Abstract |
1.Culture of Schwann Cells
We have cultured the dorsal root ganglia of rat. First, we observed the neurite outgrowth, and confirmed the following: (1)Adding the nerve growth factor over certain density brings neurite outgrowth under control all the more. (2)As a substrate, Basement Membrane MATTIGEL tm or the combination of Poly-L-Lisine and Laminine is suitable. (3)Fetal bovine serum is enough adequate, and it doesn't necessarily have to be human placental serum. It was difficult to increase the number of Schwann cells, Hile possible to obtain those cells. Further, we couldn't avoid the attaching contamination of fibroblasts.
We have also tried to culture Schwann cells by means of sciatic nerves which was more simple histologically. At that time, for the purpose of supporting Schwann cells to proliferate, we have applied bovine pituitary extract and cholera toxin; however, it hasn't led us to verify the evident effect yet because of the inappropriation of density at their application. We cope with the contamination of fibroblasts by the following methods: (1)Immunocytolysis(Monoclonal Thy-1. 1 antibody and rabbit complement). (2)Application of Cytosine arabinoside. (3)Mechanical injury to fibroblasts. It seems difficult to eliminate 100% of the contamination of fibroblasts.
Our methods to verify the existance of Schwann cells are as follow: (1)Morphological confirmation by Phase-contrast microscope. (2)Confirmation by electron microscope. (3) Confirmation by Immunofluorescence Staining.
2.Replantation of Schwann cells
We are planning to plant Schwann cells stuffed into a gel-like-substance to a living body. We are now putting in practive the preliminary experiment with only this substance stuffed in silicone tube.
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