圆背角无齿蚌血细胞培养

HEMOCULTURE OF ANODONTA WOODIANA PACIFICA

  • 摘要: 用新设计的培养基培养了圆背角无齿蚌的血细胞,在倒置相差显微镜下进行了活体观察及扫描电镜摄影.发现培养的血细胞有无颗粒细胞、颗粒细胞、透明细胞和类淋巴细胞.前三者均能伸出长的伪足和突起,与瓶壁紧密贴附,呈体外培养的成纤维细胞型;后者呈圆形,不与瓶壁贴附;四者的比例约为4:2:3:1.在活体内注射或在培养基上加入PHA和ConA,培养2-7d中,每天取部分供加入秋水仙素,用空气干燥法制片,作染色体观察,但未观察到转化细胞和有丝分裂相.研究结果表明,圆背角无齿蚌的颗粒细胞、无颗粒细胞和透明细胞在体外贴附玻璃表面的特征与高等动物的巨噬细胞类似.而不贴瓶的圆形细胞与高等动物的淋巴细胞类似.但在体外培养均不能繁殖,它们可能是高度分化的细胞.

     

    Abstract: Using the newly designed culture medium for the cell and tissue culture of freshwater bivalves, the hemolymph from the oyster, Anodont woodiana pacifica(Heude), has been cultured in vitro. Four different kinds of hemocytes in the cultured hemolymph can be observed with phase contrast microscopy:agranulocytes, granulocytes, hyalinoctes and lymphoidocytes. Agranulocytes, granulocytes and hyalinocytes are fibroblast-like and can form long pseudlopods and extensions and adhere to the culture bottle wall tightly. Lymphoidocytes are always round and never adhere to the bottle wall. The proportion of the four kinds of hemocyte is 4:2:3:1. PHA and ConA were injected into the oyster before withdrawing the hemolymph, or these molecules were added to the culture medium of hemolymph, and the hemolymph were cultured for 2-7 days. Every day, part of the cultured hemolymph was taken out for the treatment of colchicine and hypotonic solution, and the observation of mitotic chromosomes by air-drying technique. However, no mitotic figure was observed. It can be concluded that the in vitro adhering properties of granulocytes, agranulocytes and hyalinocytes of the freshwater pearl oyster. Anodont woodiana pacifica, are the same as those of macrophages of the higher animal, and lymphoidocytes of the freshwater oyster, which never adhere to the bottle, morphologically like lymphocytes of the higher animal. All of the four kinds of hemocytes cannot proliferate when cultured in vitro, which is probably because they are highly differentiated cells.

     

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