A cryopreservation solution for kidney organoids, preparation method and use method

By diluting the concentration of dimethyl sulfoxide in the frozen storage solution of kidney organoids and using KOSR serum surrogate, the problems of ice crystal formation and cytotoxicity during the frozen storage are solved, and the success rate and survival rate after frozen storage are improved.

CN119054683BActive Publication Date: 2025-03-21LEAD BIOTECHNOLOGY (KUNSHAN) CO LTD
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Patent Information

Application Number
CN202411580214.0
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Filing Date
2024-11-07
Publication Date
2025-03-21
Estimated Expiration
2044-11-07

AI Technical Summary

Technical Problem

The prior art has structural damage caused by ice crystal formation and dissolution during the freezing of kidney organoids. The high concentration of dimethyl sulfoxide is potentially toxic to cells, and the effect of serum on the differentiation of kidney organoids is uncertain, affecting the success rate of resuscitation after freezing.

Method used

A frozen storage solution is provided, including CS10 cell frozen storage solution, KOSR serum substitute and improved culture medium, dilute the concentration of dimethyl sulfoxide to achieve the lowest concentration, reduce ice crystal formation and cytotoxicity, and avoid the effect of serum on the differentiation of kidney organoids.

Benefits of technology

By diluting the concentration of dimethyl sulfoxide and using KOSR serum surrogate, ice crystal formation and cytotoxicity were reduced, and the success rate and survival rate of resuscitation after frozen renal organoids were improved.

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Abstract

The present invention provides a cryopreservation solution, a preparation method and a use method for kidney organoids, and relates to the field of cell and tissue engineering technology. The cryopreservation solution includes the following components by weight: 4.7 to 5.3 parts of cell cryopreservation solution; 3.5 to 4.5 parts of KOSR serum substitute; 0.8 to 1.2 parts of improved culture medium; wherein the improved culture medium includes RPMI 1640 culture medium and L-alanyl-L-glutamine, and the weight of RPMI 1640 culture medium is 3.5 to 4.5 times that of L-alanyl-L-glutamine. KOSR serum substitute and improved culture medium are added to CS10 cell cryopreservation solution to dilute the concentration of dimethyl sulfoxide, so that the dimethyl sulfoxide in the cryopreservation solution reaches the required minimum concentration, which not only reduces the formation of ice crystals, but also avoids the cytotoxicity caused by the excessively high concentration of dimethyl sulfoxide, thereby reducing the damage of kidney organoids.
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