| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1989: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1988: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1987: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Research Abstract |
Prior to study the transfection of genes of special materials such as hormone into goat mammary epithelial cells, it is necessary to obtain the immortal proliferation of cell and genomic DNA or cDNA of specific materials. In the present study, first, the possibility of culture of the goat mammary epithelial cells derived from goat milk was studied and the cell morphology was investigated. Second, culture of these cells in the special circumstances such as using collagen-gel or, third, condition of co-culture without cellular contact between fibroblast, were studied. Concerning about preparation of gene, on the other hand, genomic DNA derived from equine testis was analyzed by using probes of human calcitonin and calcitonin gene related peptide genes.
Results of these experiments were as follows. Condition of culture of goat mammary epithelial cells derived from milk required 30% fetal calf serum instead of 10 % at ordinal culture method. Although cell morphology changed from various types in primary culture to elongated or pleomorphic in passaged one, electron microscopical and immunohistochemical methods proved the evidence of epithelial nature of these cells. By using collagen-gel which provide similar environment as in vivo, goat mammary epithelial cells derived from milk represented round shape similar to that of in vivo, however, no growth of the cells were detected in the condition of embedding the cells in collagen-gel. Concerning about relationship between goat mammary epithelial cells and mammary fibroblasts in vitro, substituting of 50 % conditioned medium of fibroblast increased the proliferation of mammary epithelial cells. However, this was still worse than the co-culture with fibroblast. On the other hand, although calcitonin gene and calcitonin gene related peptide gene were uniformly conserved between thoroughbred breed horses, structural differences between horse and human was suggested in the present study.
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