CN102051344A - Human osteosarcoma cell line group and mouse in-vivo transplantation model - Google Patents
Human osteosarcoma cell line group and mouse in-vivo transplantation model Download PDFInfo
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Abstract
The invention discloses three human osteosarcoma cell lines well2, well 5 and LZJ, the collection numbers of the three human osteosarcoma cell lines are CCTCC C200961, CCTCC C200962 and CCTCC C200963 respectively, and the invention further discloses establishment of a corresponding NOD/SCID (nonobese diabetic/severe combined immunodeficiency disease) mouse in-vivo transplantation model, thereby providing a good and stable experimental subject and a model platform for clinical and fundamental research work of osteosarcoma. The established osteosarcoma cell lines can enable the in-vitro stable passage to be more than two years, have stable characters and enable morphology, have immuno-phenotype, chromosome genetics and other characteristics similar to the common osteosarcoma cell lines. The tumorigenesis of NOD/SCID mice is stable, the pathomorphism of tumor bodies is similar to primary tumor tissues of patients, and the animal model is simultaneously sensitive to common chemotherapeutic drugs for the osteosarcoma and can well simulate clinical retracking for clinical patients with the osteosarcoma.
Description
Technical field
The present invention relates to technical field of microbe cell line, more particularly, relate to the foundation of transplantation model in lineup's human osteosarcoma cell system and the mouse body.
Background technology
Osteogenic sarcoma is common bone malignant tumour, sends out in the teenager grade malignancy height well.Treatment plan at present commonly used is operative treatment+new adjuvant chemotherapy, and the radical cure effect is not very good, and lung shifts the incidence height.Fundamental cause is that pathogenetic understanding it be unclear that for osteosarcoma, and carrying out also of basic research work do not goed deep into.The skeletonization tumor cell line becomes the knurl transplantation model with animal be the basic tool that arrives commonly used in tumor invasion mechanism and medicine and other therapy screening study work.Though Quan Wei cell bank ATCC has had several human osteosarcoma cell systems (MG-63, Saos-2 and U-2os etc.) in the world, the tumorigenesis rate is all extremely low in animal body for they; Greatly influenced the quality of basic research work.In addition, minority Individual testwas chamber may have human osteosarcoma cell self-built, that the tumorigenesis ability is arranged and be, but there is no widely-used.Therefore, setting up the interior transplantation model of stablizing of novel human osteosarcoma cell system and body is a basic premise condition that promotes clinical and basic research work.
At present, there are MG-63, Saos-2 and U-2os etc. in home and abroad human osteosarcoma cell system commonly used.These clones are for a long time at subculture in vitro separately, and some has lost some features of the tumor tissues of original separation, especially they the tumorigenesis ability is extremely low in animal body, can not set up research model in the body.A few experiments chamber human osteosarcoma cell self-built, that the tumorigenesis ability can be arranged system, the proterties that goes down to posterity for a long time instability, be not used widely, thereby caused the restriction that becomes the knurl modelling in the osteogenic sarcoma animal body, the fundamental research of osteogenic sarcoma pathogenesis and medicine and other therapies screening operation can not be carried out in a deep going way.
Summary of the invention
The purpose of this invention is to provide three human osteosarcoma cell systems.
It is transplantation model in the NOD/SCID mouse body that second purpose of the present invention provides corresponding people's human osteosarcoma cell.
People's human osteosarcoma cell system disclosed by the invention (well5, well2 and LZJ) inoculates and the tissue derived thing that goes down to posterity in NOD/SCID mouse body from Chinese's osteogenic sarcoma tissue sample, thereafter again external stable going down to posterity more than 2 years, biological character is similar to the former foster human osteosarcoma cell of being commissioned to train with phenotype, and the tumorigenesis ability is strong on the NOD/SCID mouse.Its transplantation model tumorigenesis in NOD/SCID mouse body is stable, and the transplanted tumor pathomorphism of generation is similar to osteogenic sarcoma patient sample of former generation.The body inner model has well been simulated the process of clinical generation of osteogenic sarcoma and development.
The present invention improves and optimizes the treating processes of tissue sample, guarantees the histocyte vigor as far as possible.At first set up transplantation model in the NOD/SCID body, behind interior generation repeatedly, begin external building again and be.Carry out strict screening operation, in setting up about 19 primary cultures, filter out the clone that goes down to posterity inside and outside 3 liptinites.The human osteosarcoma cell system (well5, well2 and LZJ) that sets up is external stable going down to posterity more than 2 years, and proterties is stable, and features such as form, immunophenotype and chromosome genetics are similar to the human osteosarcoma cell of using always.Characteristics such as in addition, the human osteosarcoma cell cording of foundation has the tumorigenesis ability strong, and growth vigor is good.The NOD/SCID mouse becomes knurl stable, and knurl body pathomorphism is similar to patient's tumor tissues of former generation, and this animal model has well been simulated the osteosarcomatous clinical transition of clinical patient to osteogenic sarcoma chemotherapy medicament sensitive commonly used simultaneously.
Description of drawings
Below in conjunction with Figure of description, the present invention is described in more detail.
Figure 1A: the osteogenic sarcoma patient preceding knurl body general appearance figure that performs the operation; Figure 1B: osteogenic sarcoma patient diagnoses the CT image.
Fig. 2 A: patient specimen is digested to be injected into the NOD/SCID mouse behind the single cell suspension subcutaneous, successfully sets up and stride the species transplantation model, shown in knurl body volume bigger, be about 30mm; Fig. 2 B: after the cervical vertebra dislocation method is put to death mouse, dissect and take out the knurl piece, it is cut into fine grained chippings after, be digested to single cell suspension, be seeded to substratum, carry out cell cultures.
Fig. 3: long-term external stable cultured cells is: the light microscopic phase difference image (4X, 20X) of well2, well5, LZJ, big and sturdy, the fusiformis of cellular form is similar to human osteosarcoma cell commonly used.
Fig. 4 A: the well5 human osteosarcoma cell of foundation is that karyomit(e) quantity is unusual, and distortion obviously; Fig. 4 B: the LZJ human osteosarcoma cell of foundation is that karyomit(e) quantity is unusual, and distortion obviously.
Fig. 5: under the condition of serum-free culture, well2, well5, LZJ have respectively can be in the ability of external sphere-forming.
Fig. 6: shown in patient sample LZJ of former generation P0 similar with the pathomorphism that the NOD/SCID mouse strides species transplantation model LZJ P1, the fine osteogenic sarcoma that must reflect of this model is in the intravital generation of patient, development.
Embodiment
People's human osteosarcoma cell disclosed in this invention is, be deposited in Wuhan: Chinese typical culture collection center (CCTCC), the address: Wuhan University, preservation date is on October 29th, 2009, its name and deposit number are respectively: (1) called after: people's human osteosarcoma cell is well2; Deposit number: CCTCC-C200961; (2) name is well5 for people's human osteosarcoma cell; Deposit number: CCTCC-C200962; (3) namer's human osteosarcoma cell is LZJ; Deposit number: CCTCC-C200963.
I, mainly collect former generation sample of just sending out the osteogenic sarcoma patient in the period of the 07-08 of Rui Jin hospital, remove necrotic tissue and clot, shred the sarcoma tissue, become single cell suspension with collagenase digesting, a part is added to Tissue Culture Dish with nutrient solution after resuspended and carries out vitro culture, the direct subcutaneous implantation NOD/SCID mouse of another part treats that tumour forms.
II, observation NOD/SCID mouse become the knurl situation, and the knurl body of transplanting the back generation is carried out analyses such as pathology and immunophenotype.The tumour transplatation thing is prepared single cell suspension, go down to posterity, remove dead cell and heteroproteose cell in external continuous long-term stability.
(the well2:CCTCC-C200961 of people's human osteosarcoma cell system of III, foundation; Well5:CCTCC-C200962; LZJ:CCTCC-C200963) external stable going down to posterity more than 2 years, big and sturdy, the fusiformis of cellular form is similar to human osteosarcoma cell commonly used, and can be formed into the osteosarcoma cell ball under external serum free culture system.
Claims (6)
1. people's human osteosarcoma cell is that its deposit number is CCTCC-C200961.
2. the described human osteoblast cell of claim 1 is a transplantation model in the NOD/SCID mouse body.
3. people's human osteosarcoma cell is that its deposit number is CCTCC-C200962.
4. the described human osteoblast cell of claim 3 is a transplantation model in the NOD/SCID mouse body.
5. people's human osteosarcoma cell is that its deposit number is CCTCC-C200963.
6. the described human osteoblast cell of claim 5 is a transplantation model in the NOD/SCID mouse body.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102766600A (en) * | 2012-07-06 | 2012-11-07 | 中国人民解放军第二军医大学 | Application of esophageal squamous cell carcinoma primary tumor line CH-H-2 |
| CN110402895A (en) * | 2019-09-09 | 2019-11-05 | 中南大学湘雅二医院 | A kind of construction method of osteosarcoma xenograft mouse model |
| CN110476892A (en) * | 2019-09-09 | 2019-11-22 | 中南大学湘雅二医院 | A kind of construction method of osteosarcoma xenograft mouse model and application |
| CN110476891A (en) * | 2019-09-09 | 2019-11-22 | 中南大学湘雅二医院 | A kind of construction method of synovial sarcoma xenograft mouse model and its application |
| CN110495424A (en) * | 2019-09-09 | 2019-11-26 | 中南大学湘雅二医院 | A kind of construction method of synovial sarcoma xenograft mouse model |
| CN110495422A (en) * | 2019-09-23 | 2019-11-26 | 中南大学湘雅二医院 | A kind of construction method of malignant schwannoma xenograft mouse model and its application |
| CN110999865A (en) * | 2020-01-07 | 2020-04-14 | 中国科学院深圳先进技术研究院 | Construction method and application of osteoporosis mouse model caused by secondary hyperthyroidism |
| CN113424799A (en) * | 2021-06-28 | 2021-09-24 | 中国人民解放军陆军特色医学中心 | Construction method and application of PDX model based on osteogenic niche microenvironment modification |
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| CN110106150B (en) * | 2019-05-30 | 2020-08-11 | 中南大学湘雅二医院 | Preparation method and application of synovial sarcoma cell line hSS-005R |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102766600A (en) * | 2012-07-06 | 2012-11-07 | 中国人民解放军第二军医大学 | Application of esophageal squamous cell carcinoma primary tumor line CH-H-2 |
| CN110402895A (en) * | 2019-09-09 | 2019-11-05 | 中南大学湘雅二医院 | A kind of construction method of osteosarcoma xenograft mouse model |
| CN110476892A (en) * | 2019-09-09 | 2019-11-22 | 中南大学湘雅二医院 | A kind of construction method of osteosarcoma xenograft mouse model and application |
| CN110476891A (en) * | 2019-09-09 | 2019-11-22 | 中南大学湘雅二医院 | A kind of construction method of synovial sarcoma xenograft mouse model and its application |
| CN110495424A (en) * | 2019-09-09 | 2019-11-26 | 中南大学湘雅二医院 | A kind of construction method of synovial sarcoma xenograft mouse model |
| CN110495422A (en) * | 2019-09-23 | 2019-11-26 | 中南大学湘雅二医院 | A kind of construction method of malignant schwannoma xenograft mouse model and its application |
| CN110495422B (en) * | 2019-09-23 | 2022-12-20 | 中南大学湘雅二医院 | Construction method and application of malignant schwannoma xenograft mouse model |
| CN110999865A (en) * | 2020-01-07 | 2020-04-14 | 中国科学院深圳先进技术研究院 | Construction method and application of osteoporosis mouse model caused by secondary hyperthyroidism |
| CN113424799A (en) * | 2021-06-28 | 2021-09-24 | 中国人民解放军陆军特色医学中心 | Construction method and application of PDX model based on osteogenic niche microenvironment modification |
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| CN102051344B (en) | 2014-07-23 |
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