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SKH-1 mouse skin squamous carcinoma cell line and application thereof in preparation of transplanted tumor model

A technology of SKH-1 and cancer cells, applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc., can solve the problems of limited application, achieve good growth, fast proliferation, and make up for the effect of long modeling cycle

Active Publication Date: 2020-01-07
SHANGHAI DERMATOLOGY HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cell lines and transplantation models derived from OPN-deficient SCC tissue have certain limitations, which limit their application in the study of SCC biological characteristics and carcinogenesis mechanisms

Method used

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  • SKH-1 mouse skin squamous carcinoma cell line and application thereof in preparation of transplanted tumor model
  • SKH-1 mouse skin squamous carcinoma cell line and application thereof in preparation of transplanted tumor model
  • SKH-1 mouse skin squamous carcinoma cell line and application thereof in preparation of transplanted tumor model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] Establishment of UV-induced mouse SCC model.

[0088] Ten female SKH-1 hairless mice aged 6-8 weeks were selected, and the back skin of the mice was irradiated with a solar ultraviolet simulator close to the solar spectrum to construct a mouse skin scale model. The irradiation distance of the solar ultraviolet simulator is 30cm, and the actual output power irradiated to the mouse skin is UVB 3.0mW / cm 2 , UVA 27mW / cm 2 , the irradiation dose is the minimum erythema dose MED, 5 days a week of continuous irradiation, and after 18 weeks of continuous irradiation, new organisms > 1 mm and accompanied by ulceration can appear on the back of the mouse. The new organisms were excised by trepanning, soaked in formalin and fixed for pathological diagnosis.

[0089]24 to 48 hours after the first irradiation of the mice, erythema and fine debris appeared on the back, occasionally accompanied by slight edema. Continue to illuminate, the above skin reaction will gradually increase...

Embodiment 2

[0093] Primary culture of mouse SCC cell line.

[0094] SCC tumor-bearing mice 28 weeks after UV induction were sacrificed by cervical dislocation. Put the mouse into a dressing bowl filled with 75% alcohol and soak it for 10 minutes, cut out the tumor mass of the UV-induced SCC tumor-bearing mouse, remove the scab and necrotic tissue on the surface of the tumor, and use 2% double antibody ( 200U / ml penicillin and 200μg / ml streptomycin) were washed with PBS three times to remove the blood stains; the cleaned tumor was placed in a dressing bowl filled with high-glucose DMEM medium, and the connective tissue was cut off with ophthalmic scissors and submucosal blood vessels, cut the tumor into minced meat, about 2mm 3 Size, collected in PBS 15ml centrifuge tube containing 1% double antibody, 1000rpm / min, 5min, centrifuged and washed 3 times until the liquid was clear; discard the supernatant, add 0.25% trypsin equal to the volume of the tumor tissue block according to the size o...

Embodiment 3

[0097] Purification and passage of freshly isolated cell lines.

[0098] Fibroblasts were removed by enzymatic digestion combined with differential attachment method, and SCC cells were purified; when the cells reached 80% to 90% confluence, the cells were digested with 0.25% trypsin-EDTA trypsin and DMEM complete medium with 10% FBS After the digestion was terminated, the cells were collected by centrifugation and seeded to 25 cm 2 Inoculated culture flasks with a cell density of 3×10 5 / ml. The culture medium of SCC cells within 5 passages is DMEM medium containing 20% ​​FBS, 2% double antibodies (200U / ml penicillin and 200μg / ml streptomycin) and 10ug / ml amphotericin B, 1:2 passaging , subcultured approximately every 3 days. After the 5th generation, the culture medium of SCC cells was replaced with DMEM culture medium of 10% FBS, 1% double antibody (100 U / ml penicillin and 100 μg / ml streptomycin) and 10 ug / ml amphotericin B, and passaged according to 1:3. After 10 gener...

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Abstract

The invention discloses a SKH-1 mouse skin squamous carcinoma cell line and application thereof in the preparation of a transplanted tumor model. The SKH-1 mouse skin squamous carcinoma cell line is named as XL50 and is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC NO:C201827. The content of an obtaining method of the SKH-1 mouse skin squamous carcinoma cell line includes the steps that a SCC model is constructed by UV-induced mice; primary culture of mouse skin squamous carcinoma cell XL50 is performed; tumorigenicity test of the mouse skin squamous carcinoma cell XL50 is performed; and the SKH-1 mouse transplanted skin squamous carcinoma cell model is constructed. The cell is good in growth and can be continuously passaged and stably proliferated, the culture conditions, passage and cryopreservation methods of the cell are all conventional cell culture techniques, and the operation is convenient. The cell is fast in cell proliferationand high in degree of malignancy. The prepared cell line can be well applied to the construction of an immunogenic transplanted tumor model, not only makes up for the limitation of long modeling period of an original mouse SCC model and the tumor size, but also makes up for the lack of immune function of transplanted tumors in nude mice, and provides a good model for future research on SCC-related immune mechanisms.

Description

technical field [0001] The invention relates to a SKH-1 mouse skin squamous cell carcinoma cell line and its application in a transplanted tumor model. Background technique [0002] Skin squamous cell carcinoma (Squamous Cell Carcinoma, SCC) is one of the common human skin malignancies, accounting for about 20% of non-melanoma skin tumors. The occurrence of SCC is closely related to ultraviolet rays (Ultraviolet, UV). In recent years, as the global environmental damage has caused the damage to the ozone layer, the incidence of SCC has also increased year by year. Due to the rapid development, metastasis and poor prognosis of SCC, SCC has become the main skin tumor that threatens human health, and has attracted more and more attention. SCC is mainly related to DNA damage caused by excessive UV exposure, but its specific occurrence and development mechanism has not been fully elucidated. Traditional treatment methods, including surgery, laser, cryotherapy, and radiotherapy, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09A01K67/027C12R1/91
CPCA01K67/0271A01K2207/12A01K2227/105A01K2267/0331C12N5/0693
Inventor 王秀丽王宏伟吉杰
Owner SHANGHAI DERMATOLOGY HOSPITAL
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