ROCK Method for efficient long-term cryopreservation of intestine organoid by a combination of organoid state and optimal treatment timing of Rho-kinase inhibitor

摘要

The present invention provides an effective long-term freezing and cryopreservation method of a long orgalloid through a combined treatment of the long-augaroid state and the ROCK inhibitor addition timing. According to the present invention, it was confirmed that in vitro cultured intestinal organoids were frozen and cryopreserved in the form of intact intestine organoid than single cells. In the case of freezing frozen for more than 3 months, it was found that ROCK (Rho-kinase) inhibitor was directly added to the frozen medium and survival recovery rate was higher than pretreatment and thawing treatment. In addition, it was observed that the morphological characteristics of the in vivo intestine with the crypt-villi structure remained intact after the thawing. Therefore, the high-efficiency freezing and cryopreservation method applied to the present invention can be applied to the treatment of effective intestinal diseases and damage by providing a large amount of cells essential for organoid-based therapy.