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Cell sorting method

A cell and sorting technology, applied in the field of cell sorting, can solve the problem of blank stem cell reversible sorting, and achieve the effect of reducing the chance of pollution, simple method and low cost

Inactive Publication Date: 2015-12-23
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reversible sorting of cells refers to a cell sorting method that can release cells from the sorting carrier. In recent years, although there have been reports of reversible sorting of cancer cells using aptamers and thermosensitive polymers, but regarding stem cells reversible sorting is still blank

Method used

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  • Cell sorting method

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Embodiment

[0035] 1. Biotinylated antibodies

[0036] Dilute the CD105 antibody to be biotinylated to 1 mg / ml with 0.1 mol / L PBS buffer; dissolve NHS-biotin with DMSO to prepare a 10 mM solution, and calculate the amount of NHS-biotin required to label the antibody according to the following formula :

[0037] The amount of biotin required = the volume of the antibody × the concentration of the antibody / the molar mass of the antibody × the multiple of biotin that needs to be labeled

[0038] The antibody was mixed with NHS-biotin according to the calculated amount, and the excess biotin was removed with a 30KD ultrafiltration tube after incubation at room temperature for 30 min.

[0039] 2. Streptavidin-labeled magnetic beads

[0040] Alginic acid was activated with EDC / NHS for half an hour and incubated with streptavidin for 4 hours at 4°C. The magnetic beads are mixed with alginic acid and then added dropwise to the calcium chloride solution to form a cross-linking of alginic acid...

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Abstract

The invention provides a cell sorting method, a self-assembly system of sodium alga acid and a biotin-streptavidin system are utilized, cell sorting is conducted through combination of an antibody and specificity of a cell surface marker, and objective cells without magnetic beads are finally obtained; besides, in the sorting process, the magnetic beads are not in direct contact with the cells, and the cytotoxicity of the magnetic beads and mechanical damage to the cells are avoided. The method includes the steps that marking biotin through the antibody, marking the magnetic beads through the streptavidin, incubating the antibody marked by the biotin and the cells, incubating the cell-antibody compound and the magnetic beads marked by the streptavidin, conducting magnetic gathering on cell-magnetic bead compound and disintegrating the cell-magnetic bead compound and obtaining the objective cells with the magnetic beads. According to the cell sorting method, the self-assembly system of sodium alga acid, a biotin-streptavidin system and an antigen-antibody system are combined, the advantages of magnetic bead sorting can be given play, the cell sorting experiment is simpler in process, more cells are obtained, and the quality is higher.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a cell sorting method. Background technique [0002] With the development of cell replacement therapy, stem cell transplantation therapy has become an important means of clinical treatment of certain diseases. The use of stem cells expanded and cultured in vitro and induced into the required cells is then transplanted into the patient's body for the repair of tissue damage, Replacement of degenerative organs and improvement of function of genetically defective tissues and organs. The discovery of plastic differentiation of adult stem cells provides a new direction for cell replacement therapy. Despite a number of advances in the field of cell therapy, significant challenges remain. Achieving the goal of treating a disease requires obtaining the precise population of cells needed to treat the target disease and, once transplanted, ensuring that these cells reach the site whe...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 曾令文林姣
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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