Method of separating cattle marrow mesenchyme stem cell

Abstract

the invention discloses a separating method of ox marrow mesenchyma stem cell, which comprises the following steps: adding ox marrow single-cell suspension liquid in the Percoll cell separating liquid with density at 1.082 to separate; obtaining foggy single-core cell at liquid intersecting layer; inoculating in the plastic container; removing non-adhesive wall cell to obtain the product with separating purifying efficiency over 98%.

Description

technical field [0001] The invention relates to a method for isolating bovine bone marrow mesenchymal stem cells. Background technique [0002] With the rapid development of life sciences, the improvement of local varieties has become more and more simple, resulting in the loss of many local characteristic varieties and the emergence of new varieties. Therefore, the establishment of variety gene pools is very important. Bovine bone marrow mesenchymal stem cells are one of the seed cells of the gene bank of fine breeds of cattle, and bone marrow mesenchymal stem cells have certain medical application value. Bone marrow mesenchymal stem cells (mesenchymal stem cells, MSCs), originally discovered by Friedenstein, are a type of cells that are easy to attach to the surface of plastic culture plates. Later studies have confirmed that under different induction conditions, they have mesoderm and neuroectoderm tissues. The ability of cell differentiation, such as the ability to diff...

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Problems solved by technology

[0003] It has been reported that MSCs are difficult to isolate and are often contaminated by hematopoietic progenitor cells

Commonly used MSCs isolation and culture methods are: 1. Density gradient centrifugation: according to the density difference between MSCs and other cells, Percoll layering solution is used to separate MSCs, but the acquisition rate of MSCs is very low; 2. Adherence screening method: According to the characteristics of MSCs that grow on the wall in plastic tissue culture flasks, it is generally advisable to use 4-12 hours to adhere to the cells. Although the separation of MSCs is very uniform at first, the colony formation rate is low; 3. Flow cytometry Sorting method: According to the small size of MSCs cells and the relative lack of particles, MSCs are sorted by flow cytometry, but the confocal laser has a toxic effect on the activity of cells; Enrichment of certain expressed cell surface markers or negative sorting based on negatively expressed antigens on the surface has high efficiency and purity, but targeting MSCs-specific surface antigens is expensive, complicated to operate, and difficult to sort

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