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Mice animal model of giant cell tumor of bone of human and construction method thereof

A giant cell tumor of bone and giant cell tumor technology, which is applied in the field of human giant cell tumor mouse animal model and its construction, achieves the effect of simple technical operation, unique construction method and high success rate

Inactive Publication Date: 2014-07-09
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, there is currently no literature report on the successful establishment of an animal model of human giant cell tumor of bone.

Method used

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  • Mice animal model of giant cell tumor of bone of human and construction method thereof
  • Mice animal model of giant cell tumor of bone of human and construction method thereof
  • Mice animal model of giant cell tumor of bone of human and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1: Construction of mouse animal model of human giant cell tumor bone injury of the present invention

[0037] 1. Isolation and culture of primary giant cell tumor cells

[0038] Surgical specimens diagnosed as giant cell tumor of bone by clinical tissue biopsy were placed in normal saline, blood clots, necrotic tissue, fat and connective tissue on the surface of the tumor tissue were removed under aseptic conditions, and the tissue was cut into pieces with sterilized ophthalmic scissors. Wash 2-3 times with sterile PBS, add a small amount of PBS, use elbow ophthalmic scissors, and cut the tissue repeatedly until the tissue becomes a paste, about 1mm 3 size. Add type II collagenase, digest at 37°C for 30 minutes, add 5ml of α-MEM medium containing 10% FBS to stop digestion, filter through a 100um filter, centrifuge at 800rmp for 5min, remove the supernatant, add 10ml of α-MEM containing 10%FBS Resuspend the cells in the culture medium, inoculate the cell sus...

Embodiment 2

[0042] Example 2: Study on the characteristics of the mouse animal model of human giant cell tumor of bone obtained by the construction method in Example 1

[0043] Example 1 Two months after the injection of primary giant cell tumor cells into the tibia, the nude mice were anesthetized and sacrificed, the tibia was taken out and fixed with 4% paraformaldehyde for 24 hours, decalcified with 0.5M EDTA for 10 days, and then treated with graded ethanol , Xylene, soaked in paraffin (that is, dehydrated to wax treatment), embedded in paraffin, cut into 4uM thick slices. HE, TRAP and anti-human mitochondrial immunofluorescence staining were used to observe the gross morphological changes, osteoclast activity and cell source of nude mouse tibia.

[0044] 1. The characteristics of osteolytic lesions in the tibia of nude mice injected with primary cells of human giant cell tumor of bone are consistent with the clinical X-ray manifestations of giant cell tumor of bone, that is, simple o...

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Abstract

The invention relates to an animal model and a construction method thereof; the incidence rate of giant cell tumor of bone (GCT) is high in China, and an animal model of giant cell tumor of bone of human is not successfully established. The invention provides a mice animal model of giant cell tumor of bone of human and a construction method thereof. Primary cells obtained by isolated culture of a giant cell tumor tissue of bone of human are injected into shin bones of nude mice to obtain the mice animal model of giant cell tumor of bone of human. Osteolytic lesions features of the mice animal model of bone injury of giant cell tumor of bone of human are similar to pathological features of clinical giant cell tumor of bone, and the survival cells are human-derived stromal cells of giant cell tumor of bone. The mice animal model of giant cell tumor of bone of human can be sued for systematic study on pathogenesis of the giant cell tumor of bone and further screening of drugs for treatment of the giant cell tumor of bone.

Description

technical field [0001] The invention relates to an animal model and a construction method thereof, in particular to a human giant cell tumor mouse animal model and a construction method thereof. Background technique [0002] Giant cell tumor of bone (GCT) has a high incidence in my country, accounting for about 20% of all bone tumors, and the spine is one of the predilection sites for GCT (see literature Sung H.W., Kuo D.P., Shu W.P. , et al. Giant-cell tumor of bone: analysis of two hundred and eight cases in Chinese patients [J]. J Bone Joint Surg Am, 1982,64(5):755-761.). The Orthopedic Oncology Department of Changzheng Hospital has treated an average of more than 60 patients with spinal GCT every year in the past five years. GCT is highly invasive and highly recurrent, and the postoperative local recurrence rate is as high as 30-50% (see literature Hart R.A., Boriani S., Biagini R., et al.A system for surgical staging and management of spine tumors. A clinical outcome s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09A01K67/02A61K49/00
Inventor 肖建如徐乐勤吴志鹏杨兴海周振华黄权林在俊万维矫健蔡小攀陈素许炜祝倾刘铁龙严望军杨诚钱明李珍惜周旺
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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