CAJ | 학술논문

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25~30 g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100 mm3。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。
목적：탐토이용다발성골수류환자골수류세포경식입토골건립 NOD/SCID 소서다발성골수류모형적가행성。방법분리취출신서란백토고골여경골,청제기육、골막급연골조직,연중간경유절단；준비4~6주체질량약25~30 g적 NOD/SCID 소서,복강주사마취,장토골식입소서후배부,4주후검측토골식입성활정황。장분선적다발성골수류환자 CD38+/CD45-적장세포5×106경과면역형광표기후,유원측단완완주입식입토골내。매주관찰성류정황,채용 living-Imaging 방법검측소서체내장세포적생장정황,병리검측종괴성질。결과토골식입후4주성활,장분선적다발성골수류환자골수류세포주입토골후,2주가견 NOD/SCID 소서하류,8주후종류체적대소약100 mm3。living-Imaging방법검측결과현시,재주사후2주가견토골국부유장세포농취,수착주사시간적연장,지주사후8주국부장세포농취현상경가명현(2.4×104 vs .1.5×105,P <0.05)。종괴병리활검가견대량장세포침윤,식입토골결구파배,파골세포증다。결론NOD/SCID 소서토골식입가유효지지다발성골수류환자장세포국부주사,병건립 NOD/SCID 소서골수류모형,위골수류상관체내실험급림상약물사선제공실험모형。
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 - 30 g (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 100 mm3 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 (2.4×10 4 vs .1.5× 10 5 ,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.