模式原生动物浮萍棘尾虫纯培养体系的建立与优化

浮萍棘尾虫(Stylonychia lemnae)是一种营动物性营养的单细胞真核生物, 是研究细胞学、遗传学问题的重要模式原生动物, 而实现浮萍棘尾虫纯培养是开展棘尾虫生物学研究的重要基础。研究以建立高效棘尾虫无菌纯培养体系为目的, 参考嗜热四膜虫无菌纯培养基, 采用响应面分析方法, 对氮源、碳源和磷酸氢二钾的组分配比进行单因素实验和正交实验, 初步建立用于浮萍棘尾虫无菌培养的培养基, 并分别对培养温度、初始pH、培养液添加量和起始接种密度等培养条件进行了优化。结果表明: 起始接种量为100 cells, 培养基10 mL, 初始pH 7.0, 温度25℃, 静置培养168h, 可获得最大细胞数量约为3.0×103 cells。

ESTABLISHMENT AND OPTIMIZATION OF PURE CULTURE SYSTEM OF A MODEL PROTOZOA, STYLONYCHIA LEMNAE

Stylonychia lemnae is a monocyte eukaryote with animal nutrition. It is also an important model protozoa to study cytology and genetics. Sterile pure culture is an important basis for the study of Stylonychia biology. In order to build a high-efficiency pure cultured system for Stylonychia, a Tetrahymena thermophila sterile pure cultured medium was referenced in this paper. And a response surface analysis was used to carried out single-factor and orthogonal experiments focused on the ratio of nitrogen source, carbon source and dipotassium hydrogen phosphate. A sterile pure cultured medium for Stylonychia was primarily established and some key cultured conditions were optimized including cultured temperature, initial pH, medium volume and initial cell density. Results showed that the securable maximum cell amount was about 3.0×103 cells and the best conditions were initial amount 100 cells, medium volume 10 mL, initial pH 7.0 and the cultured temperature 25℃.