4results about How to "Short doubling time" patented technology

The invention discloses a serum substitute compatible umbilical cord mesenchymal stem cell culture medium additive composition and application thereof. The serum substitute compatible umbilical cord mesenchymal stem cell culture medium additive composition is prepared from the following components in parts by mass: 15 to 25 parts of platelet-derived growth factor-BB; 15 to 25 parts of basic fibroblast growth factors; 8 to 12 parts of transforming growth factor-beta; and 8-12 parts of recombinant human insulin. The serum substitute compatible umbilical cord mesenchymal stem cell culture medium additive composition and the application thereof disclosed by the invention have the following beneficial effects: (1) the proliferation efficiency is remarkably improved; (2) the cell morphology and dryness can be well maintained; and (3) the compliance and the compatibility are good.

The application discloses a method for inducing T cell reprogramming into NK-like cells, comprising the following steps: (1) culturing the T cells in a culture system to obtain activated T cells; (2) adding small molecule inducers twice to the activated T cells to obtain NK-like cells; wherein the culture system in step (1) contains 1-10 v / v % serum substitute and / or platelet lysate, and does not contain fetal bovine serum. By optimizing the culture process and precisely adding drugs twice, the small molecule accumulation toxicity caused by multiple drug additions is avoided, the damage to the structure and function of the mitochondrial membrane is reduced, the mitochondrial membrane potential is significantly improved, the energy metabolism capacity of the mitochondria is enhanced, sufficient energy support is provided for the long-term survival and function maintenance of the cITNK cells, and the physiological activity of the cells is ensured from the energy metabolism level.

The application discloses a kind of human mesenchymal stem cell 3D culture and amplification method, belong to stem cell culture amplification technical field.The application is prepared 3D cell culture matrix by structure design and component optimization, realize the accurate simulation of stem cell in-vivo microenvironment.The matrix is modified sodium alginate by amino hexene acid and is compounded with methacryl gelatin, and is crosslinked by lithium chloride and calcium chloride, and is synergistically formed by photocrosslinking, and the three-dimensional network has uniform pore size and excellent connectivity.The application significantly improves cell adhesion rate and proliferation efficiency, effectively maintains the biological characteristics of hMSC such as immune regulation and differentiation, the degradation rate of the matrix is dynamically matched with the cell proliferation cycle, the material has excellent biocompatibility and the degradation product is non-toxic, which solves the defects of traditional 2D culture and existing 3D technology, and provides an efficient and safe technical solution for hMSC scale-up and clinical translation.

The present application relates to a kind of preparation method for preparing neural stem cell with high proliferative capacity, belong to the field of biotechnology.The method is isolated from mammalian embryo brain tissue primary neural stem cell, with DMEM / F12 as base medium, add B27, N2, glutamine and double resistance, then join epidermal growth factor, basic fibroblast growth factor, insulin-like growth factor-1, Y-27632 and heparin form optimized proliferation culture medium.Cell is seeded in the culture dish coated with polylysine and laminin, culture under suitable conditions, neural ball reaches certain diameter after digestion and passage.This method can improve the proliferation efficiency of neural stem cell, maintain stemness and differentiation potential, adopt serum-free system, simple operation, suitable for large-scale expansion and research application.