水生生物学报
水生生物學報
수생생물학보
ACTA HYDROBIOLOGICA SINICA
2001年
2期
116-122
,共7页
石安静%邱安东%唐敏%余燕萍%张洪渊%町井昭
石安靜%邱安東%唐敏%餘燕萍%張洪淵%町井昭
석안정%구안동%당민%여연평%장홍연%정정소
圆背角无齿蚌%血细胞%培养
圓揹角無齒蚌%血細胞%培養
원배각무치방%혈세포%배양
Anodonta woodiana pacifica%Hemocyte%Culture
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。
用新設計的培養基培養瞭圓揹角無齒蚌的血細胞,在倒置相差顯微鏡下進行瞭活體觀察及掃描電鏡攝影。髮現培養的血細胞有無顆粒細胞、顆粒細胞、透明細胞和類淋巴細胞。前三者均能伸齣長的偽足和突起,與瓶壁緊密貼附,呈體外培養的成纖維細胞型;後者呈圓形,不與瓶壁貼附;四者的比例約為4∶2∶3∶1。在活體內註射或在培養基上加入PHA和ConA,培養2—7d中,每天取部分供加入鞦水仙素,用空氣榦燥法製片,作染色體觀察,但未觀察到轉化細胞和有絲分裂相。研究結果錶明,圓揹角無齒蚌的顆粒細胞、無顆粒細胞和透明細胞在體外貼附玻璃錶麵的特徵與高等動物的巨噬細胞類似。而不貼瓶的圓形細胞與高等動物的淋巴細胞類似。但在體外培養均不能繁殖,它們可能是高度分化的細胞。
용신설계적배양기배양료원배각무치방적혈세포,재도치상차현미경하진행료활체관찰급소묘전경섭영。발현배양적혈세포유무과립세포、과립세포、투명세포화류림파세포。전삼자균능신출장적위족화돌기,여병벽긴밀첩부,정체외배양적성섬유세포형;후자정원형,불여병벽첩부;사자적비례약위4∶2∶3∶1。재활체내주사혹재배양기상가입PHA화ConA,배양2—7d중,매천취부분공가입추수선소,용공기간조법제편,작염색체관찰,단미관찰도전화세포화유사분렬상。연구결과표명,원배각무치방적과립세포、무과립세포화투명세포재체외첩부파리표면적특정여고등동물적거서세포유사。이불첩병적원형세포여고등동물적림파세포유사。단재체외배양균불능번식,타문가능시고도분화적세포。
Using the newly designed culture medium for the cell and tissueculture 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. Anodonta 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.