小鼠单核巨噬细胞白血病细胞(RAW264.7)的培养及其在诱导破骨细胞中的应用

Culture skill of RAW264.7 and its application in osteoclastic differentiation

小鼠单核巨噬细胞白血病细胞(RAW264.7)是通过Abelson鼠白血病病毒诱导BALB/c小鼠产生肿瘤后得到的细胞株。RAW264.7细胞在医学研究中应用十分普遍:它是许多白血病相关研究模型的常用细胞株;在微生物学、免疫学等研究中也十分常用;同时也是小鼠源性破骨前体细胞,可通过特定细胞因子的诱导在体外获得成熟破骨细胞,因此也成为许多骨骼疾病研究的体外模型。但RAW264.7细胞形态和分化状态多变,在实际培养中较难把握,给科研工作带来了一定困难。科研工作者在RAW264.7细胞的培养方法、细胞的形态及分化状态等方面始终观点不一,对于用RAW264.7细胞在体外诱导获得破骨细胞的方法也有许多差别。本文结合文献资料及实践,探索了RAW264.7细胞的培养条件,冻存、复苏、传代方法,以及用核因子κB受体活化因子配基(receptor activator for nuclear factor-κB ligand,RANKL)诱导其成为成熟破骨细胞的技术关键;旨在总结RAW264.7细胞的培养及诱导其分化为破骨细胞的经验教训,探讨RAW264.7细胞的培养和在体外用RANKL诱导RAW264.7细胞分化为破骨细胞的方法、技巧,供广大科研工作者借鉴。

The RAW264. 7 cell line was obtained from a tumor of BALB / c mouse induced by Abelson murine leukemia virus.RAW264. 7 cell was broadly applied in medical sciences： it is the mostly adopted cell line in many leukemia-related research. It is also commonly used in immunity and microbiology research. In addition,it is the murine source pre-osteoclast which could be stimulated with cytokines and differentiate into mature osteoclasts in vitro,thus becoming an in vitro model applied in many skeletal diseases. However,its cellular morphology and differentiation stage are variable and hard to handle and therefore brings some difficulties to practical work. As to the methods of induction of RAW264. 7 cells to mature osteoclasts,researchers opinions are also different. In this article,we explored the culture condition with combination of literature study,figured out the methods of cryopreservation,recovery and subculture of RAW264. 7 cells,and stimulated it with the receptor activator for nuclear factor-κB ligand（ RANKL）. The aims of this article are to summarize experience of culturing RAW264. 7 cell and its differentiation into mature osteoclast,and to explore the related skills,thus providing reference to researchers.