Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Preparation method for immortalized lymphocytes

A lymphocyte and immortalization technology, which is applied in the field of preparation of immortalized lymphocytes, can solve the problems of cumbersome preparation methods and low success rate, and achieve the effects of wide host range, high success rate and improved sensitivity

Inactive Publication Date: 2013-09-25
HARBIN MEDICAL UNIVERSITY
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, immortalized lymphocytes are obtained by Epstein-Barr virus infection, transfection of oncogene or telomerase, the preparation method is cumbersome and the success rate is low

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Step 1: Immunizing mice with specific antigens.

[0029] When implementing it:

[0030] Select four-week-old BAL B / C mice to immunize the mice. According to the different characteristics of the antigen, it is divided into the following two schemes:

[0031] (1) Soluble antigen: The immunogenicity is weak, and adjuvants are generally added. For haptens, immunogens should be prepared first, and then adjuvants should be added. Commonly used adjuvants: Freund's complete adjuvant, Freund's incomplete adjuvant.

[0032] Initial immunization: 1-50 μg antigen plus Freund's complete adjuvant subcutaneous multi-point injection or intrasplenic injection (generally 0.8-1ml, 0.2ml / point)

[0033] After 3 weeks, the second immunization: the dose is the same as above, plus Freund’s incomplete adjuvant subcutaneously or ip (intraperitoneal injection) (ip dose should not exceed 0.5ml)

[0034] After 3 weeks, the third immunization: the same dose, no adjuvant, ip (5 to 7 days later, b...

Embodiment 2

[0053] Step 1: Immunizing mice with specific antigens. (step 1 with embodiment 1)

[0054] Step 2: Obtain lymphocytes from the spleen.

[0055] When implementing it:

[0056] (1) Three days after the booster immunization, the mice were killed by dislocation of the neck and disinfected by immersing in 75% ethanol. Take out the spleen of the mouse in the ultra-clean bench, grind the spleen with 4-5ml 1× lymphocyte separation medium (shake well before taking it) in a sterile grinding tube, and filter it with a 200-mesh sieve after grinding to ensure that the isolated cells for a single cell suspension.

[0057] (2) Immediately transfer the separation liquid containing the spleen cells to a 15ml centrifuge tube, cover with 200-500 μL of 1640 medium, and centrifuge at 800 g for 30 min at room temperature.

[0058] (3) Aspirate the lymphocyte layer, add 10ml of 1640 medium, and wash upside down. Cells were collected by centrifugation at 250 g for 10 min at room temperature. Po...

Embodiment 3

[0065] Step 1: select mice naturally infected with leukemia virus, and take spleen under aseptic conditions.

[0066] Step 2: obtaining lymphocytes from the spleen, culturing the lymphocytes, and obtaining immortalized lymphocytes.

[0067] When implementing it:

[0068] (1) Grind the spleen with 4-5ml 1× lymphocyte separation medium (shake well before use) in a sterile grinding tube, and filter it with a 200-mesh sieve after grinding to ensure that the isolated cells are a single cell suspension.

[0069] (2) Immediately transfer the separation liquid containing the spleen cells to a 15ml centrifuge tube, cover with 200-500 μL of 1640 medium, and centrifuge at 800 g for 30 min at room temperature.

[0070] (3) Aspirate the lymphocyte layer, add 10ml of 1640 medium, and wash upside down. Cells were collected by centrifugation at 250 g for 10 min at room temperature. Pour off the supernatant, resuspend the cells with lymphocyte culture medium, and culture them with soft agar...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a preparation method for immortalized lymphocytes. Lymphocytes are obtained by infecting animal spleens with leukemia virus of the same animal; the lymphocytes which are infected by leukemia virus are cultivated; and then the immortalized lymphocytes are obtained.

Description

technical field [0001] The invention relates to a preparation method of immortalized lymphocytes. Background technique [0002] After being induced by a specific antigen, immortalized lymphocytes can produce specific antibodies or cytokines against the antigen. In the prior art, immortalized lymphocytes are obtained by Epstein-Barr virus infection, transfection of oncogene or telomerase, the preparation method is cumbersome and the success rate is low. Contents of the invention [0003] The object of the present invention is to provide a method for preparing immortalized lymphocytes, which is simple and easy to operate, and has a high success rate. [0004] The technical scheme that realizes the object of the present invention: [0005] A preparation method of immortalized lymphocytes, characterized in that: after immunizing animals, the lymphocytes produced in the spleen of the same animal are infected with animal leukemia virus, and the aforementioned lymphocytes infec...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N5/078
Inventor 张凤民李玉军宋武琦李辉吴静
Owner HARBIN MEDICAL UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com