日本七鳃鳗外周血细胞显微结构及类淋巴细胞体外培养

以日本七鳃鳗(Lampetra japonica)为研究对象,观察其外周血细胞显微结构和探讨类淋巴细胞原代培养条件。采用Wright氏和Giemsa氏染色法对日本七鳃鳗外周血液有形成分进行显微观察,可鉴别出红细胞、淋巴细胞、中性粒细胞、单核细胞、嗜酸性粒细胞;未发现嗜碱性粒细胞。通过Ficoll密度梯度离心法和流式细胞仪分选获得高纯度的七鳃鳗类淋巴细胞。经条件优化后确定最适培养液L15+20%FBS(5%七鳃鳗血清)+0.46%NaCl,在18℃,pH 6.8-7.0的条件下对日本七鳃鳗类淋巴细胞进行体外培养,大多数类淋巴细胞半贴壁生长,细胞状态较好,最长可以存活近一个月。类淋巴细胞的原代培养为日本七鳃鳗细胞体外培养的深入研究提供了实验依据。

Microscopic structures of Japanese lamprey peripheral blood cells and culture of lymphocyte-like cells in vitro

The microstructure of Japanese lamprey(Lampetra japonica) peripheral blood cells and the cultivation methods of lymphocyte-like cells were explored.The peripheral blood cells of L.japonica were stained by Wright’s and Giemsa’s staining methods and five major cell types including erythrocyte,lymphocyte,neutrophil granulocyte,monocyte and eosinophil granulocyte,but not basophil were recognized under light microscope.The lymphocyte-like cells were separated successfully with Ficoll-Paque centrifugation and flow cytometry and were cultured in L15 medium with 20% foetal bovine serum(FBS) including 5% L.japonica FBS and 0.46% NaCl.The optimal temperature of cells is 18 ℃ and the optimal pH of cells is 6.8-7.0.They can develop attachment as a sheet of cells after 1 hour.Cells have a good condition for nearly a month.The establishment of primary cell cultures of lymphocyte-like cells from L.japonica provides a useful tool for the longevity of the culture and the investigation in cellular biology.