Pei Yu1, Junxiang Wen1, Jun Wang2, Jing Liang2, Yuhui Shen1, Weibin Zhang1. 1. Department of Orthopedics, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China. 2. Shanghai Institute of Traumatology and Orthopedics, Shanghai 200025, China.
Abstract
BACKGROUND: A large number of osteosarcoma patients are dying of pulmonary metastasis in spite of the recent progress achieved in treatment research. Therefore, there is an urgent need for an in vivo experiment, whose purpose is to investigate the growth and metastasis of human osteosarcoma tumor. In this study, a novel human osteosarcoma cell line was established and characterized for osteosarcoma metastatic research in vivo. METHODS: Small pieces of parental primary osteosarcoma samples from a 16-year-old boy were xeno-transplanted into NOD/SCID mice. Then, osteosarcoma cells from the xeno-grafts were isolated and further passaged in vitro. Expression profiling was confirmed using HE staining and immunohistochemistry. The tumorigenic and metastatic ability of the established cell line was analyzed by cell scratch and CCK8 assay in vitro, and tumor transplantation in NOD/SCID mice. RESULTS: Parental cells were pleomorphic and positive with ALP, SSEA-4, and CD44. Osteosarcoma cells, named Well5 cells, were successfully isolated and had the ability of adipogenesis and osteogenesis. Well5 cells were mostly positive for SSEA-4 protein, and possessed morphological characteristics such as osteoblastic nature during the cultivation and hetero transplantation. Cell scratch and CCK8 assay indicated Well5 cells obtained a better capacity for migration and proliferation than the MG63 cell line. In NOD/SCID mice, orthotopic tumors were established at the primary site, and spontaneous pulmonary metastases were developed. CONCLUSIONS: A novel human osteosarcoma cell line was successfully established, which may be helpful for spontaneous pulmonary metastasis research in vivo. 2019 Annals of Translational Medicine. All rights reserved.
BACKGROUND: A large number of osteosarcoma patients are dying of pulmonary metastasis in spite of the recent progress achieved in treatment research. Therefore, there is an urgent need for an in vivo experiment, whose purpose is to investigate the growth and metastasis of human osteosarcoma tumor. In this study, a novel human osteosarcoma cell line was established and characterized for osteosarcoma metastatic research in vivo. METHODS: Small pieces of parental primary osteosarcoma samples from a 16-year-old boy were xeno-transplanted into NOD/SCID mice. Then, osteosarcoma cells from the xeno-grafts were isolated and further passaged in vitro. Expression profiling was confirmed using HE staining and immunohistochemistry. The tumorigenic and metastatic ability of the established cell line was analyzed by cell scratch and CCK8 assay in vitro, and tumor transplantation in NOD/SCID mice. RESULTS: Parental cells were pleomorphic and positive with ALP, SSEA-4, and CD44. Osteosarcoma cells, named Well5 cells, were successfully isolated and had the ability of adipogenesis and osteogenesis. Well5 cells were mostly positive for SSEA-4 protein, and possessed morphological characteristics such as osteoblastic nature during the cultivation and hetero transplantation. Cell scratch and CCK8 assay indicated Well5 cells obtained a better capacity for migration and proliferation than the MG63 cell line. In NOD/SCID mice, orthotopic tumors were established at the primary site, and spontaneous pulmonary metastases were developed. CONCLUSIONS: A novel human osteosarcoma cell line was successfully established, which may be helpful for spontaneous pulmonary metastasis research in vivo. 2019 Annals of Translational Medicine. All rights reserved.
Entities:
Keywords:
Human osteosarcoma; Well5 cell line; spontaneous pulmonary metastasis; xeno-transplantation
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