| Culture of spermatogenic cells in vitro is an important means in procreate develop. At present, by means of factitious hormones regulation study on spermatogenesis in vitro has come true the process of proliferation and differentiation of spermatoginum, and the process of meiosis from spermatocyte to spermatid in models of mammalia, amphibia and fish. But mature of the spermatid was still hard to achieved. Establishing model of culture of spermatogenic cells in vitro is help to studying the regulate mechanism of spermatogenesis. So far, report on spermatogenesis in vitro in crustacean was lacking. Crustacean has independence and consummate regulate system of incretion early, realizing the effect of incretion and nerve regulate system on procreate throughly has important referrence significance on exploring the procreate regulate mechanism of atitude life-form.In addition, environment factor and poisonous substance has grave effect on idioplasm especialy, the idioplasm is worse and worse, restrict the develop of marine lives. Studing effects of poisonous substance on reproduce and differentiation of spermatogenic cells has theoretics significance on clarifing the mechanism of poisonous substance, and has practice significance on idioplasm profect and health breed aquatics.In this paper, spermatogenic cells in testis of Macrobrachium nipponense as a research objective, effects of 17a-MT, cadmium and androgenic gland on differentiation of spermatogenic cells from M. nipponense in vitro was studied. These results were following:(1)Under cultured spermatogenic cells with sertoli cells, the different concentrations of hormone which was supplemented to the culture medium with 17a-MT were evaluated the spermatogenic cells percentage motility, minority of spermatocytes occurred meiosis to form a tetrad of spermatids and minority of spermatids developed into spinous, it is concluded that 17a-MT has no effect on the percentage motility of spermatogenic cells in vitro, the concentration of 1500 ng/ml 17a-MT supplemented to culture medium is the suitable hormone concentration for culture spermatogenic cells in vitro.(2)Spermatogenic cells was isolated and sublimated witn trypsin digestion method and differential speed adherence, and was disposed with the different concentrations of cadmium which was supplemented to the culture medium, then was evaluated the growth restrain with MTT, DNA damage with single cell gel electrophoresis assay, we found①Spermatogenic cells was obtained more using the concentration of trypsin digestion of 0.05% and 40 minute, somatic cells and spermatids can be wiped off with differential speed adherence method.②After spermatogenic cells was disposed with cadmium,24 hours ago, spermatogenic cells had not distinct reproduce restrain, after 24 hours, concentration of 5μg/L of spermatogenic cells had not distinct reproduce restrain,50μg/L,500μg/L and 1000μg/L had distinct reproduce restrain, the absorbency fell followed the concentration of cadmium heighten.③Concentration of 5μg/L,50μg/L,500μg/L and 1000μg/L of spermatogenic cells can induce DNA damage which can be evaluated at the time of 24 hour, degree of DNA damage and all the damaged cells augment followed the concentration of cadmium heighten at the same time; degree of DNA damage and all the damaged cells augment followed the time prolonged under the same concentration. Degree of DNA damage and all the damaged cells was direct correlation with the concentration and time.(3)Experiment of spermatogenic cells cultured with androgenic gland showed:minority of spermatids developed into spinous and acrosomal vesicle was more than the control group, the result indicated that androgenic gland can promote the differentiation of spermatids but had no effect on the percentage motility of spermatogenic cells, can not improve the percentage motility of spermatogenic cells.
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