Human umbilical cord mesenchymal stem cell culture method

Abstract

The invention discloses a human umbilical cord mesenchymal stem cell culture method, which comprises: (1) obtaining an umbilical cord tissue; (2) separating Wharton's Jelly; (3) cutting the Wharton's Jelly into tissue blocks, inoculating the tissue blocks into a T75 culture bottle, adding an appropriate amount of a primary culture medium, placing into a CO2 incubator, carrying out standing culture for 3 days, supplementing 5 ml of the primary culture medium at the 4th day, changing the culture medium every 2-3 days from the 4th day, removing the tissue block when cells start to migrate from the majority of the tissue blocks, changing the culture medium, and carrying out passage amplification when the cell density achieves 80% and above; and (4) after the treatment, re-suspending with a passage culture medium. According to the present invention, the animal-derived additive is not used during the culture process, such that the safety is provided; and the operation is simple and convenient, and the culture time is short.

Description

technical field [0001] The invention relates to a method for culturing cells, in particular to a method for culturing human umbilical cord mesenchymal stem cells, and belongs to the technical field of cell culture. Background technique [0002] Mesenchymal Stem Cells (MSCs) are derived from mesoderm and ectoderm in the early stages of development and exist in connective tissues and organ tissues throughout the body. Adult stem cells with multilineage differentiation potential. [0003] MSCs can differentiate into various tissue cells such as fat, bone, cartilage, muscle, tendon, ligament, nerve, liver, heart muscle and endothelium under specific induction conditions in vivo or in vitro, and they are still multidirectional after continuous subculture and cryopreservation. Therefore, it can be used as an ideal seed cell for the repair of tissue and organ damage caused by aging and disease. [0004] MSCs can secrete and produce a variety of cytokines that promote growth and d...

AI Technical Summary

Problems solved by technology

[0009] Both the enzymatic hydrolysis method and the tissue block culture method introduce components derived from animals. Since the components such as enzymes and serum derived from animals are not clear, the risk of microbial contamination such as viruses, fungi, and mycoplasma is increased. Serum cannot be excluded for at least one year The volatile substances contained in the serum may change the normal state of cells in the body when used in serum, which poses a safety hazard to the human body

[0010] In addition, due to differences between animal-derived enzymes and serum batches, which will also reduce the reproducibility of cell culture, both enzymatic hydrolysis and tissue block culture methods have problems such as complicated culture operations and long culture times

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