CD4 + T monoclonal cell culture medium, culture method and application of CD4 + T monoclonal cell culture medium

By using a culture medium formulation of N-acetylcysteine ​​and Trolox and a gradient dilution method, the CCR5-Δ32CD4+T monoclonal cell line was successfully constructed, solving the problem that natural CD4+T cells are difficult to form monoclonal lines, improving gene editing efficiency, and providing a new method for the treatment of HIV-related diseases.

CN120924490APending Publication Date: 2025-11-11WUHAN BODA BIOTECHNOLOGY CO LTD
0 Cites 0 Cited by

Patent Information

Application Number
CN202511061253.4
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-07-30
Publication Date
2025-11-11

AI Technical Summary

Technical Problem

Existing technologies make it difficult to directly form stable natural CD4+ T cell monoclonal lines, and traditional CRISPR/CAS9 editing polyclonal cell lines have low editing efficiency, resulting in poor clinical treatment effects.

Method used

A CD4+ T monoclonal cell culture system was constructed using a culture medium formulation containing N-acetylcysteine ​​and Trolox, combined with gradient dilution and density gradient dilution methods. The CCR5-Δ32 monoclonal cell line was obtained through gene editing.

Benefits of technology

This study achieved efficient amplification of monoclonal cell lines with uniform genetic background, improving the efficiency of gene-edited cell line preparation and providing important scientific research reference for the treatment of CCR5 gene-related diseases.

✦ Generated by Eureka AI based on patent content.
Patent Text Reader

Abstract

The invention provides a CD4 + T monoclonal cell culture medium, a culture method and application of the CD4 + T monoclonal cell culture medium. The culture medium comprises N-acetylcysteine and Trolox. According to the invention, the long-standing technical problem that natural CD4 + T cells are difficult to directly and stably form a monoclonal line is successfully solved. Through a density gradient dilution method and a culture medium formula, the formation of a monoclonal cell line with uniform and stable genetic background by efficient amplification starting from a single CD4 + T cell is realized for the first time. The CD4 + T cell is high in monoclonal formation efficiency and good in monoclonal growth state, and the preparation efficiency of the gene editing cell strain is greatly improved. According to the invention, a cell line with hCCR5-delta 32 CD4 + T cell point mutation is successfully constructed by using a monoclonal cell culture method, and important scientific research reference data is provided for treatment of diseases related to the CCR5 gene.
Need to check novelty before this filing date? Find Prior Art