Three-dimensional single cell source cell ball production chip, preparation method thereof and application thereof

Abstract

The invention discloses a three-dimensional single cell source cell ball production chip, including a substrate and a pattern microarray formed on a surface of the substrate, wherein the pattern microarray includes a plurality of patterns spaced apart from each other, the substrate surface is electrically neutral, and the pattern is formed of a positively charged material. The invention also discloses a preparation method and an application of the chip. For example, one method of producing a single cell-derived mesenchymal stem cell sphere based on the chip comprises using the chip to perform3-5 days of three-dimensional culture, the spheres could be diluted by phosphate buffer and collected by centrifugation. The chip of the invention has the advantages of simple structure, lower cost and easy fabrication, and the single cell source mesenchymal stem cell ball production method based on the chip can screen and produce single cell source cell balls with high quality and homogenization,has good reliability, high stability and high throughput, and can well meet the requirements of basic research and clinical application.

Description

technical field [0001] The present invention relates to a method for producing mesenchymal stem cell spheroids, in particular to a three-dimensional single cell-derived cell spheroid production chip, its preparation method and application, such as a method for producing mesenchymal stem cell spheroids under three-dimensional culture conditions and its use in Use in promoting angiogenesis in vivo. Background technique [0002] Mesenchymal stem cells (MSC, mesenchymal stem cells) are derived from the mesoderm in the early stage of development. MSCs were first discovered in bone marrow, and later research confirmed that MSCs widely exist in various tissues and organs: including bone marrow, fat, umbilical cord blood, amniotic membrane, thymus, dental pulp, peripheral blood, umbilical cord, etc. Under specific induction conditions in vivo or in vitro, mesenchymal stem cells can differentiate into fat, bone, cartilage, muscle, tendon, ligament, nerve, liver, cardiac muscle, endo...

AI Technical Summary

Problems solved by technology

[0007] 2) There are differences in the metabolism of nutrients, oxygen and waste inside and outside the multicellular sphere. It is difficult for the cells in the center of the sphere to obtain sufficient nutrients and oxygen, and it is difficult for the waste produced by it to metabolize to the environment in a timely manner. In this process, this stress affects the gene expression level of the spheroid cells, and may even cause gene mutations, thereby affecting the quality of the cells

[0008] 3) The cells in the center of the MCS sphere are prone to anoikis due to the lack of extracellular matrix support

[0009] 4) The multicellular spheroids formed by the existing 3D culture method are generally larger in diameter, ranging from several hundred microns to several millimeters. Due to the risk of blood vessel blockage, larger-sized MCS should not be directly injected into the body through blood vessels, and high-intensity enzymatic hydrolysis Processing enables the recovery of single cells from multicellular spheroids, but the process severely damages the cells, resulting in large heterogeneity in the quality of single cells obtained

[0010] 5) The existing technology of forming cell spheroids is basically suspension culture, without the participation of extracellular matrix, it is difficult to simulate the growth environment of MSC in the real body

Images