A method for recovering and culturing ROBO1 CAR NK cells

By using HIPP-T009 lymphocyte serum-free culture medium and WAVE wave bioreactor to optimize the resuscitation and amplification culture method of CAR NK cells, the problem of small scale and high cost of CAR NK cell culture in the prior art is solved, and high-efficiency and low-cost large-scale cell production is achieved.

CN116254233BActive Publication Date: 2025-08-05SICHUAN ASCLEPIUS BIOTECHNOLOGY CO LTD +1
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Patent Information

Application Number
CN202211724587.1
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Filing Date
2022-12-30
Publication Date
2025-08-05
Estimated Expiration
2042-12-30

AI Technical Summary

Technical Problem

The existing CAR NK cell culture methods are complex in operation, limited in scale, high in cost and large batch variations, which limit their large-scale promotion and clinical application in tumor treatment.

Method used

Resuscitation culture was performed using HIPP-T009 lymphocyte serum-free medium containing IL2 and nicotinamide, and primary and secondary amplification culture was carried out in combination with WAVE wave bioreactor. The cell seeding density and processing interval time were controlled, and the culture conditions were optimized to maintain cell activity and amplification scale.

Benefits of technology

The rapid resuscitation and large-scale culture of CAR NK cells were achieved, which reduced production costs, improved cell viability and killing activity, and met the clinical use needs of allogeneic patients.

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Abstract

The present application relates to the fields of bioengineering and technology, and specifically to a method for resuscitating and culturing ROBO1CAR NK cells. The method comprises: resuscitating and culturing the cells using a resuscitation medium, wherein the resuscitation medium is a serum-free HIPP-T009 lymphocyte culture medium containing IL-2 and nicotinamide; performing a primary amplification culture on the resuscitated cells using a first amplification culture medium to obtain primary amplified cells; and performing a secondary amplification culture on the primary amplified cells using a second amplification culture medium to harvest the cells. The method can rapidly resuscitate frozen cells, effectively maintain cell growth and cell killing activity, and expand the culture scale, greatly reducing the culture cycle and production costs.
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