Immortal human ovary carcinoma cell strain and its establishment method

A method and technology for establishing ovarian cancer, applied in the field of immortalized cell lines of human ovarian sarcoma-like carcinoma, to achieve the effects of high line formation efficiency, high malignancy, and enrichment of in vitro cell banks

Inactive Publication Date: 2004-02-25
PEOPLES HOSPITAL PEKING UNIV
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Problems solved by technology

Therefore, it is extremely necessary to establish cell lines of corresponding pathological types for in-depth research in vitro, but there is no report on the establishment of a line at home and abroad.

Method used

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  • Immortal human ovary carcinoma cell strain and its establishment method
  • Immortal human ovary carcinoma cell strain and its establishment method
  • Immortal human ovary carcinoma cell strain and its establishment method

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Embodiment Construction

[0038] One, the construction of SV40 gene eukaryotic expression vector

[0039] pS 3 neo contains SV40 T antigen (including large T and small T antigen) gene fragments, which can be digested with BamH I to obtain a 3kb SV40 T gene fragment, which is recovered by electrophoresis. pcD2 is a high-efficiency expression eukaryotic vector plasmid, which is digested with BamH I at the multiple cloning site, and the linear vector is recovered by electrophoresis, and then treated with calf intestinal alkaline phosphatase to prevent self-ligation. Ligate it with the recovered SV40 T gene fragment under the action of T4 DNA ligase to obtain the recombinant plasmid pcD2-SV40. Recombinant plasmids were identified by enzyme digestion. The positive clone of pcD2-SV40 was screened out by SmaI and HpaI double digestion. The pSV 3 Both neo and pcD2 were donated by Dr. Li Huiping, Institute of Hematology, Peking University People's Hospital.

[0040] 2. Primary culture of human ovarian sarc...

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Abstract

The present invention relates to the method of establishing human ovary cancer immortal cell strian, the immortal gene-SV40T antigen gene is introduced into human ovary cancer original generation cell, then screened, resistance clone enlarge culture to obtain immortal cell strain. The established ovary cancer cell strain is named as BVPH:OVSC-2 and is kept by CGMCC with keeping No.0606, its biological characteristics includes strong invading powder, cell is spindle sarcoma call shape, apparent heteromorphism, retains biological characters of original cell.

Description

Technical field: [0001] The invention belongs to the field of microbial animal cell lines, and in particular relates to a method for establishing an immortalized human ovarian cancer cell line and an immortalized human ovarian sarcoma-like carcinoma cell line established according to the method. Background technique: [0002] Among the malignant tumors of the female reproductive system, the incidence of ovarian cancer ranks second, but the mortality rate ranks first. Despite continuous improvements in surgery, chemotherapy, and radiation, the five-year survival rate for ovarian cancer has hovered around 30 percent for the past decade. The main reason is that the pathological types of ovarian cancer are complicated, the degree of malignancy is high, the individual differences are large, and the mechanism of its occurrence and development at the molecular and cellular levels is unknown. Since the lifespan of fresh cancer cells obtained from patients is too short, it is imposs...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/16C12N15/09C12N15/85
Inventor 冯捷崔恒刘广芝钱和年付天云叶雪
Owner PEOPLES HOSPITAL PEKING UNIV
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