Adult nephroblastoma HANB cell strain and culturing method and application thereof

A technology for Wilms tumor and a culture method, which is applied to the field of adult Wilms tumor cell line and its culture, can solve the problems of difficulty in researching the mechanism of drug sensitivity and other problems, and achieves the effects of rapid growth and simple culture.

Inactive Publication Date: 2012-07-11
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no immortalized cell line of Wilms tumor, which brings difficulties to its in vitro therapeutic experimental research, drug sensitivity research and the research on the mechanism of occurrence.

Method used

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  • Adult nephroblastoma HANB cell strain and culturing method and application thereof
  • Adult nephroblastoma HANB cell strain and culturing method and application thereof
  • Adult nephroblastoma HANB cell strain and culturing method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Adult nephroblastoma HANB cell line, the preservation number is CCTCC NO: C201133, and its culture method is as follows:

[0039] 1. Culture and passage

[0040] Centrifuge the postoperative ascites from a patient with poorly differentiated anaplastic Wilms tumor at 1000 rpm for 10 minutes, discard the supernatant, and wash the precipitate with 1640 culture medium to obtain primary cells. 1640 culture medium is prepared from 1640 medium Cheng, 1640 culture medium is a commodity sold on the market, produced by American Life Tech Company, and sold by Beijing Dongge Biotechnology Co., Ltd.; the preparation method of 1640 culture medium is: 10g 1640 medium, 5.0g HEPES, 3.7g NaHCO 31. Dissolve 3g of L-glutamine in 1L of deionized water, filter through a 0.22μm microporous filter, sterilize and subpackage, and refrigerate at 4°C. On the ultra-clean workbench, place the primary cells in a cell culture flask, suspend them in 1640 culture medium with 20% newborn calf serum, and...

Embodiment 2

[0048] Adult nephroblastoma HANB cell line, the preservation number is CCTCC NO: C201133, and its culture method is as follows:

[0049] In step 1 of the culture method of adult nephroblastoma HANB cell line culture method, the primary cells are placed in culture flasks on the ultra-clean workbench, and cultured with 1640 containing 5% newborn calf serum. Liquid suspension, at 37°C, CO 2 Culture in a cell incubator with a volume concentration of 5% and a humidity of 95%, and change the 1640 culture medium containing 5% newborn calf serum every 24 hours until the cells grow to the plateau stage and complete the first stage of cell growth. For the second subculture, the cells grown to the plateau stage were subcultured; in the subculture step, digested with 0.22% trypsin aqueous solution, discarded the trypsin aqueous solution, and used newborn calf serum containing 1% mass fraction The 1640 culture medium was blown on the bottle wall to suspend the cells, and the suspension wa...

Embodiment 3

[0052] Adult nephroblastoma HANB cell line, the preservation number is CCTCC NO: C201133, and its culture method is as follows:

[0053] In step 1 of the culture method of adult nephroblastoma HANB cell line culture method, the primary cells are placed in culture flasks on the ultra-clean workbench, and cultured with 1640 containing 20% ​​newborn calf serum. Liquid suspension, at 37°C, CO 2 Culture in a cell incubator with a volume concentration of 5% and a humidity of 95%, and change the 1640 culture medium containing 15% newborn calf serum every 24 hours until the cells grow to the plateau stage and complete the first stage of cell growth. For the second subculture, the cells that have grown to the plateau stage are subcultured; in the subculture step, digest with a trypsin aqueous solution with a mass fraction of 0.28%, discard the trypsin aqueous solution, and use a newborn calf serum with a mass fraction of 5% The 1640 culture medium was blown on the bottle wall to suspe...

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Abstract

The invention discloses an adult nephroblastoma HANB cell strain with the collection number of CCTCC NO:C201133. A culturing method comprises the following steps of: culturing in-vitro ascites of a patient suffering from poorly-differentiated anaplastic nephroblastoma; passing; freezing for preserving; and recovering; the cell growing speed is high, the quantity is large, the quality is uniform, and a cell culturing method is simple; as proved by evaluation on the biological characteristics such as cell form, protein structure, chromosome, neoplastic rate and the like of the cell strain, the cell strain keeps the biological characteristics of a primary tumor; and due to the usage of the adult nephroblastoma HANB cell strain to the establishment of a nephroblastoma model, a research model is provided for the occurrence, transfer mechanism and clinical treatment of the nephroblastoma.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an adult nephroblastoma cell line and a culture method thereof. Background technique [0002] Wilms' tumor or Nephroblastoma, also known as Wilms' tumor, occurs in 90% of children before the age of 6, and its incidence rate ranks first among abdominal tumors in children. It is a common abdominal solid malignant tumor. It originates from primitive Wilms tissue and can form an embryonic structure. It is rare in adults and is not sensitive to radiotherapy and chemotherapy. It is highly malignant and prone to distant metastasis, often with a poor prognosis. Cell tumors have important clinical significance. However, there is currently no immortalized cell line of Wilms tumor, which has brought difficulties to its in vitro therapeutic experimental research, drug sensitivity research and pathogenesis research. [0003] Therefore, obtaining Wilms tumor cell lines with immortali...

Claims

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Application Information

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IPC IPC(8): C12N5/09A01K67/027
Inventor 邵晨张磊师长宏赵清波汪涌王晓武窦小亮乔少谊
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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