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Cancer Research

Establishment of Patient-Derived Xenograft Mouse Model with Human Osteosarcoma Tissues

Published: March 22nd, 2024



1Longhua Hospital, Shanghai University of Traditional Chinese Medicine, 2Key Laboratory of theory and therapy of muscles and bones, Ministry of Education
* These authors contributed equally

The present protocol describes the method for establishing a patient-derived xenograft (PDX) mouse model using human osteosarcoma tissue.

Osteosarcoma is the most common primary malignant bone tumor in children and adolescents. Despite the development of new treatment plans in recent years, the prognosis for osteosarcoma patients has not significantly improved. Therefore, it is crucial to establish a robust preclinical model with high fidelity. The patient-derived xenograft (PDX) model faithfully preserves the genetic, epigenetic, and heterogeneous characteristics of human malignancies for each patient. Consequently, PDX models are considered authentic in vivo models for studying various cancers in transformation studies. This article presents a comprehensive protocol for creating and maintaining a PDX mouse model that accurately mirrors the morphological features of human osteosarcoma. This involves the immediate transplantation of freshly resected human osteosarcoma tissue into immunocompromised mice, followed by successive passaging. The described model serves as a platform for studying the growth, drug resistance, relapse, and metastasis of osteosarcoma. Additionally, it aids in screening the target therapeutics and establishing personalized treatment schemes.

Osteosarcoma is a primary bone malignancy derived from interosseous lobe cells and is most common in adolescents as well as children. It often occurs in the epiphysis of the long diaphysis and is characterized by high malignancy, early metastasis, and poor prognosis1,2. Lung metastasis is the main cause of death in osteosarcoma patients. The 5-year survival rate of patients with non-metastatic osteosarcoma is 65%-70%3. However, over the last 40 years, the 5-year survival rate (only 20%) of patients with metastatic osteosarcoma has not significantly improved, and 25% of osteosarcoma pati....

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All studies involving human tissues have been approved by the Institutional Ethics Review Committee of Longhua Hospital, affiliated with Shanghai University of Traditional Chinese Medicine (Shanghai, China) (2013LC52), and written informed consent was obtained from the patients in accordance with the Helsinki Declaration. The IACUC number for this animal study is PZSHUTCM221017013. Four-week-old male BALB/c thymus-free mice were provided with specific pathogen-free (SPF) feed and sterile water, and were housed in a singl.......

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This protocol describes the detailed procedure for establishing a PDX mouse model, preserving the morphological features of human osteosarcoma after immediate transplantation of freshly resected human osteosarcoma tissue and successive passages in mice. Here, a PDX mouse model was successfully established using human osteosarcoma tissue.

Figure 3A shows a representative mouse of PDX at P0, two months after the transplantation of femoral osteosarcoma tissue from a .......

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The PDX models can simulate the characteristics of human cancers and retain more similarity with the primary tumor, including genetic and genomic alterations, histology, heterogeneity, and gene expression profile16,17,18,19. Therefore, they preserve the molecular phenotypes and genotypes of cancer patients, providing innovative approaches for studying biology and evaluating potential therapeuti.......

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This work is supported by grants from (1) the National Nature Science Foundation (81973877 and 82174408); (2) Shanghai Top Priority Research Center construction project (2022ZZ01009); (3) National Key R&D Program of China (2020YFE0201600); (4) Shanghai Collaborative Innovation Center of Industrial Transformation of Hospital TCM Preparation and (5) Research Projects within Budget of Shanghai University of Traditional Chinese Medicine (2021LK047).


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NameCompanyCatalog NumberComments
10% formalin neutral solutionWuhan Saiweier Biotechnology Co., LtdG1101-500mlFix the tissues
AutoclaveJapan Hiryama CompanyHVE-50Sterilization surgical instruments
BALB/c athymic miceShanghai SLAC Laboratory Animal Co, Ltd./Animal
CaliperYantai Green Forest Tools Co., Ltd.034180AMeasure the tumor volume
Dish (60mm)Shanghai NianYue Biotechnology Co., Ltd430166, CorningSample placment during transplantation
Disinfectant cotton ballsShanghai Honglong Industrial Co., Ltd.20230627Disinfect the skin of mice
Disposable sterile glovesGuilin Hengbao Health Protection Co., Ltd.YT21131Sterile operation
Electronic scaleShanghai NianYue Biotechnology Co., Ltd1-2000Weigh the weight of the tumor
EosinShanghai Gengyun Biotechnology Co., LtdE4009-25GHematoxylin eosin stain
HematoxylinShanghai Gengyun Biotechnology Co., LtdH3136-25GHematoxylin eosin stain
IsofluraneShenzhen RWD Life Technology Co., LtdVETEASYMouse anesthesia 
Mark penZebra Trading (Shenzhen) Co., Ltd.YYST5Mark the surgical incision
Olympus Optical microscopeJapanese Olympus CompanyBH20Scan tissue slices
Ophthalmic ointmentShanghai Gengyun Biotechnology Co., LtdSOICOEYEGRLAvoid dry eyes of mice during anesthesia
Ophthalmic scissorsShanghai NianYue Biotechnology Co., LtdY00030 JZCut the skin
Ophthalmic tweezersShanghai NianYue Biotechnology Co., LtdBS-ZER-S-100 BiosharpHold osteosarcoma tissues during transplantation
ParaffinJiangsu Shitai Experimental Equipment Co., Ltd.80200-0015Buried osteosarcoma tissue
Paraffin slicing machineLyca Microsystem (Shanghai) Trading Co., Ltd.RM2235Osteosarcoma tissue section
physiological salineGuangzhou Jinsheng Biotechnology Co., Ltd.605-004057Rinse and temporary storage of osteosarcoma tissue
ScalpelsSurgical Instrument Factory of Shanghai Medical Devices (Group) Co., Ltd.J11010-10# JZSeparation of osteosarcoma tissue and making surgical incisions
Sterile hoodThermo Fisher Technology (China) Co., Ltd.ECO0.9Surgical operation table
sterile surgical drapesHenan Huayu Medical Equipment Co., Ltd.20160090Provide sterile surgery area
Straight needle holderShanghai Gengyun Biotechnology Co., LtdJ31050 JZSuture the wound
Suture lineShanghai Pudong Jinhuan Medical Products Co., LtdF3124Suture the wound
Suture needleShanghai Pudong Jinhuan Medical Products Co., LtdF3124Suture the wound
Tissue protective solutionNanjing Shenghang Biotechnology Co., LTDBC-CFM-03Maintain the activity of tissue cells
Tube (50 mL)Shanghai Baisai, Biotechnology Co., Ltd.BLD-BL2002500Install formalin fixation solution

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