A kind of composition, culture medium and application for siRNA transfection 3D organoid

Abstract

The invention provides a composition, culture medium and application for siRNA transfection of 3D organoids. The composition comprises: at least one of serum albumin and ovalbumin compounded with at least one of connexin and serum substitute. The medium includes: basal medium; at least one of the following final concentrations of extracellular matrix components to promote transfection: 1‑10% HSA (wt / v), 1‑10% BSA (wt / v), 1‑10% OVA (wt / v); transfection-promoting 3D organoid components consisting of at least one of the following final concentrations: 10‑500 ng / mL connexin‑43, 10‑500 ng / mL connexin‑26, 1‑5% serum replacement. The technology of the siRNA transfection 3D organoid of the present invention is less time-consuming, has high efficiency, does not need to mechanically destroy the shape of the 3D organoid, has a wide range of applications, and can be widely used for siRNA transfection of cancer or normal tissue organoids.

Description

technical field [0001] The invention relates to the technical field of siRNA transfection cells, in particular to a composition, culture medium and application for siRNA transfection of 3D organoids. Background technique [0002] Organoids are cell cultures with a 3D structure grown in vitro using adult stem cells, which have highly similar histological features to corresponding human organs and can reproduce the physiological functions of the organ. Tumor organoids (tumoroids) are "micro-tumors" that are cultured in vitro using patient tumor tissue and are highly consistent with the structure and physiology of the patient's tumor. Organoids can be used as in vitro models of various diseases, and have broad application prospects in stem cells and development, regenerative medicine, disease research, drug development, and precision medicine. Organoid models can replicate the tissue complexity and genetic heterogeneity of tumors. Compared with many preclinical models such as ...

AI Technical Summary

Problems solved by technology

The transfection method of siRNA in 2D cell lines has been successfully established, but in the application of 3D organoids, it is usually necessary to mechanically destroy the formed organoids and prepare them into 2D cells for transfection, which is inefficient and interferes with the growth state of organoids , but the success rate of direct application is lower

At present, some researchers are focusing on transfecting siRNA into organoids by in situ method, but this method has not seen any technical progress

Images