A method for three-dimensional cell culture using injectable thermosensitive hydrogel

By using an injectable thermosensitive hydrogel precursor solution to mix with cells and rapidly incubate, combined with a vitamin B6 derivative degradation method, the problems of excessively long phase transition time and insufficient stability of synthetic thermosensitive hydrogels in three-dimensional cell culture were solved, achieving high-quality cell culture and rapid cell recovery.

CN122303137APending Publication Date: 2026-06-30BEIHAO STEM CELL & REGENERATIVE MEDICINE RES INST CO LTD +1
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
BEIHAO STEM CELL & REGENERATIVE MEDICINE RES INST CO LTD
Filing Date
2024-12-31
Publication Date
2026-06-30

AI Technical Summary

Technical Problem

Existing synthetic thermosensitive hydrogels suffer from problems such as excessively long phase transition times and insufficient material stability in three-dimensional cell culture, leading to reduced cell activity and material loss. Furthermore, commonly used natural matrix gels have unclear composition and ethical issues.

Method used

After mixing the injectable thermosensitive hydrogel precursor solution with the cells, the mixture was incubated at 34-37℃ for 1-10 minutes, then cultured in cell culture medium. During the incubation degradation process, a vitamin B6 derivative was used for rapid degradation. After centrifugation, the cultured cells were obtained.

Benefits of technology

It achieves rapid sol-gel phase transition, provides a stable three-dimensional culture environment, ensures cell growth and differentiation under physiological conditions, and enables rapid and complete degradation of the gel structure after culture, achieving efficient cell recovery.

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Abstract

This application proposes a method for three-dimensional cell culture using an injectable thermosensitive hydrogel. Cells are mixed with a thermosensitive hydrogel precursor solution and incubated at 34-37°C for 1-10 minutes, followed by the addition of cell culture medium. After culture, a vitamin B6 derivative is added for degradation, yielding a cell suspension. The cultured cells are then separated by centrifugation. Using a hydrogel with a rapid sol-gel phase transition provides cells with a three-dimensional support environment close to the in vivo environment, enabling cells to grow, proliferate, differentiate, and interact with other cells and the matrix under similar physiological conditions, achieving high-quality in vitro cell culture. Specifically, the three-dimensional structure is less affected by temperature during culture, which is beneficial for stable cell culture. After culture, the gel structure degrades rapidly and completely, allowing for rapid cell recovery; this is a high-quality cell culture method.
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