利用患者骨髓瘤细胞建立NOD/SCID小鼠多发性骨髓瘤模型
Establishment of NOD/SCID-mouse model with multiple myeloma using myeloma cells from patients

摘要：目的 探讨利用多发性骨髓瘤患者骨髓瘤细胞经植入兔骨建立NOD/SCID小鼠多发性骨髓瘤模型的可行性。方法 分离取出新西兰白兔股骨与胫骨，清除肌肉、骨膜及软骨组织，沿中间轻柔截断；准备4～6周体质量约25～30g的NOD/SCID小鼠，腹腔注射麻醉，将兔骨植入小鼠后背部，4周后检测兔骨植入成活情况。将分选的多发性骨髓瘤患者CD38+/CD45-的浆细胞5×106经过免疫荧光标记后，由远侧端缓缓注入植入兔骨内。每周观察成瘤情况，采用living-Imaging方法检测小鼠体内浆细胞的生长情况，病理检测肿块性质。 结果 兔骨植入后4周成活，将分选的多发性骨髓瘤患者骨髓瘤细胞注入兔骨后，2周可见NOD/SCID小鼠荷瘤，8周后肿瘤体积大小约100mm3。living-Imaging方法检测结果显示，在注射后2周可见兔骨局部有浆细胞浓聚，随着注射时间的延长，至注射后8周局部浆细胞浓聚现象更加明显(2.4×104 vs. 1.5×105，P<0.05)。肿块病理活检可见大量浆细胞浸润，植入兔骨结构破坏，破骨细胞增多。结论 NOD/SCID小鼠兔骨植入可有效支持多发性骨髓瘤患者浆细胞局部注射，并建立NOD/SCID小鼠骨髓瘤模型，为骨髓瘤相关体内实验及临床药物筛选提供实验模型。
ABSTRACT: Objective To explore the feasibility of establishment of NOD/SCID-mouse model with multiple myeloma by using plasma cells from myeloma patients. Methods The femurs and tibias were removed from the New Zealand white rabbits; the muscles, periosteum and cartilage tissues were cleared. Then each bone was cut into two pieces gently along its middle. The NOD/SCID mice weighing 25-30g (4-6 weeks) were anesthetized by intraperitoneal injection; rabbit bone was inserted into the right side of the mouse back and engraftment of the bones was allowed to take place after 4 weeks. The 5000 000 purified plasma cells which expressed CD38+/CD45- were immunofluorescence labeled and then injected slowly into the implanted rabbit bone through the distal end. The mice were observed weekly; the plasma cells growth in mice was screened by the living-imaging system and the tumor from the mice was determined by biopsy. Results The implanted rabbit bone survived after 4 weeks. The tumor in mice was observed 2 weeks after the purified myeloma cells were injected into the rabbit bone, and it reached 100mm3 after 8 weeks. Results of the living-imaging system showed that the myeloma cells had uptake in the rabbit bone after 2 weeks of injection and this phenomenon was more pronounced after 8 weeks of injection (2.4×104 vs. 1.5×105, P<0.05). The tumor infiltrated with numerous plasma cells and osteoclasts increased upon the biopsy. Conclusion Rabbit bone marrow implanted into NOD/SCID mice can effectively support local injection of plasma cells of multiple myeloma patients, and the NOD/SCID-mouse model of myeloma has been established. This model can be used to study in vivo experiments related to myeloma and clinical therapeutic approaches for this disease