A Treg cell expansion and culture process

Through specific amplification culture medium formula and technical means, the activation and expansion process of Treg cells is optimized, and the problems of low proliferation capacity and rapid decrease in the proportion of Treg cells in the prior art are solved, achieving efficient and long-lasting Treg cell expansion.

CN119776277AActive Publication Date: 2025-04-08SHANGHAI SAIERXIN BIOMEDICAL TECH CO LTD
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Patent Information

Application Number
CN202510280567.7
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-03-11
Publication Date
2025-04-08
Estimated Expiration
2045-03-11

AI Technical Summary

Technical Problem

The existing Treg cell amplification methods have low proliferation ability, high frequency of cell aging or apoptosis, and limited amplification days, resulting in a rapid decrease in the proportion of CD4+CD25+CD127lo/-.

Method used

Specific amplification culture medium formulations, including AB serum, IL-2, rapamycin, α-tocopherol, elamod and gynostemophenol, combined with density gradient centrifugation and flow sorting techniques, optimize the activation and amplification process of Treg cells.

Benefits of technology

The activity and passage number of Treg cells was significantly improved, the number of cell expansion days was extended, and the proportion of CD4+CD25+CD127lo/-Treg cells was maintained, and even at 78 days still accounted for a relatively high proportion.

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Abstract

The present invention discloses a Treg cell expansion and culture process, which relates to cell therapy. The process comprises: step 1, separating umbilical cord blood to obtain a peripheral blood mononuclear cell layer; step 2, using flow cytometry to sort CD4 + CD25 + CD127 lo / ‑ Treg cells; step three, Treg cell activation and expansion, cord blood Treg cells cryopreservation; the expansion culture medium includes: AB serum, IL-2 concentration of, rapamycin concentration of, α-tocopherol, iguratimod, and gypenosides; the formula of the present invention, α-tocopherol, iguratimod, and gypenosides are synergistic in increasing the Treg ratio and cell activity; the formula and method of the present invention can prolong the expansion days, and can have a good expansion effect even at 78 days.
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