人源骨肉瘤原代细胞的分离及其CDX模型和PDX模型的构建

目的建立骨肉瘤患者人源肿瘤细胞系异种移植(CDX)模型和人源移植(PDX)模型,获得与患者肿瘤细胞相似的原代肿瘤细胞。方法获取1例右胫骨骨肉瘤患者的新鲜肿瘤活性组织,无菌条件下转移至实验室。将其中一部分剪切为2mm3组织块,种植于培养皿中。获取细胞后,使用胰蛋白酶消化法和反复贴壁法分离原代肿瘤细胞。将原代肿瘤细胞植入BALB/c裸鼠背部皮下和胫骨髓腔,待肿瘤长至直径1~2cm时,收获组织,制备HE切片。将另一部分组织植入BALB/c裸鼠背部皮下,待肿瘤体积长至800~1500mm3,取出一代肿瘤组织,将其中部分冻存,其余组织再次植入BALB/c裸鼠背部皮下传代。结果所获得CDX模型和PDX模型肿瘤细胞与患者病理切片细胞形态相似。结论采用患者骨肉瘤组织原代细胞分离与BALB/c裸鼠肿瘤移植等方法,可建立患者来源的骨肉瘤CDX和PDX模型并获得与患者肿瘤细胞相似的原代肿瘤细胞。

Isolation of primary human osteosarcoma cells and establishment of cell-derived xenograft model and patient-derived xenograft model

Objective To establish the cell-derived xenograft (CDX) model and the patient-derived xenograft (PDX) model and obtain primary tumor cells of osteosarcoma patient. Methods The fresh active tissue of an osteosarcoma patient was obtained and transferred to the laboratory under aseptic conditions. Some of the tissue was cut into 2 mm 3 size and cultured in petri dishes. After obtaining the cells, trypsin digestion and repeated adherent method were used to isolate the primary tumor cells. Primary tumor cells were implanted in the dorsal subcutaneous and tibial medullary cavity of BALB/c nude mice. When the tumor grew to 1-2 cm in diameter , tissue was harvested and HE sections were prepared. Another part of tissue was implanted the dorsal subcutaneous of BALB/c mice. When the tumor volume reached 800-1 500 mm 3, the tumor tissue was removed, part of P1 tissue was frozen, and the rest tissue was implanted the dorsal subcutaneous of nude mice again for passage . Results The primary tumor cells were polygonal and spherical. The morphology of tumor cells in CDX model and PDX model was highly comparable patterns of that in pathological sections of patients. Conclusion Primary tumor cells may be obtained by establishing CDX and PDX models of osteosarcoma patient.