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Mouse skin digestion complex enzyme and method for isolating micro-immunity Treg cells from mouse skin

A technology of immune cells and compound enzymes, applied in the medical field, can solve the problems of immature technology, low efficiency, and small number of cells, and achieve good digestion effect, good digestion effect, and convenient cytology and molecular research.

Pending Publication Date: 2019-10-08
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

[0004] In a study on the mechanism of extracting mouse skin immune cell subsets published in 2014, a method for preparing a single-cell suspension of skin tissue from mouse skin tissue was reported, but the number of cells detected was small, the technology was not mature, and In subsequent studies, the less efficient method is still used

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  • Mouse skin digestion complex enzyme and method for isolating micro-immunity Treg cells from mouse skin
  • Mouse skin digestion complex enzyme and method for isolating micro-immunity Treg cells from mouse skin
  • Mouse skin digestion complex enzyme and method for isolating micro-immunity Treg cells from mouse skin

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Embodiment Construction

[0027] The invention provides a mouse skin digestion complex enzyme and a method for isolating trace immune cells Treg cells from mouse skin, wherein: the mouse skin digestion complex enzyme is composed of RPMI (Roswell Park Memorial Institute) medium, 1% HEPES ( 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) buffer, 10% fetal bovine serum, 1% penicillin / streptomycin double antibody solution, 2mg / ml type 11 collagenase (Collegenase11), 0.5mg / ml hyaluronic acid Enzyme (hyaluronidase, HAase) and 2mg / ml deoxyribonuclease (deoxyribonuclease, DNase) prepared. Collagenase is an enzyme extracted from bacteria. It has a strong digestive effect on collagen and is suitable for digesting fibrous tissue, epithelial tissue and cancer tissue. It has little effect, and can separate cells from collagen components without damage. The addition of other enzymes can get better digestion effect.

[0028] Based on the mouse skin digestion compound enzyme provided above, the present invention ...

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Abstract

The invention belongs to the field of medical science, and relates to a mouse skin digestion complex enzyme and a method for isolating micro-immunity Treg cells from mouse skin. The mouse skin digestion complex enzyme is prepared by a RPMI medium, a HEPES buffer solution with a final concentration of 1%, fetal bovine serum with the final concentration of 10%, a penicillin / streptomycin double-antibody solution with the final concentration of 1%, type 11 collagenase with the final concentration of 2 mg / ml, hyaluronidase with the final concentration of 0.5 mg / ml, and deoxyribonuclease with the final concentration of 2 mg / ml. The method can efficiently obtain the micro-immunity Treg cells from a limited size mouse skin tissue, which facilitates cytological and molecular studies on skin-specific Treg cells, and also provides a reference for separating other micro cells in the skin. .

Description

technical field [0001] The invention belongs to the medical field, and relates to a digestive complex enzyme and a cell separation method based on the digestive enzyme, in particular to a mouse skin digestive complex enzyme and a method for isolating trace immune cells Treg cells from mouse skin. Background technique [0002] Skin diseases such as contact dermatitis, eczema, psoriasis, cellulitis, fungal infections and abscesses, and non-melanoma skin cancers are among the 50 most common diseases worldwide, research and elucidate the molecular and cellular mechanisms of different skin diseases is an essential avenue for the development of effective targeted therapeutic strategies. Mouse models are an important tool for studying the occurrence and development of diseases and targeted therapy, especially in the field of skin disease research. Efficient isolation of trace cell subpopulations in the skin is a technical problem that needs to be solved urgently. [0003] Regulato...

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Application Information

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IPC IPC(8): C12N9/16C12N9/26C12N9/64C12N5/0783
CPCC12N9/6416C12N9/16C12N9/2474C12Y302/01035C12Y301/21001C12N5/0637C12N5/0636C12N2509/00C12N2501/734C12N2501/73
Inventor 李春英郭森倪清蓉李舒丽易秀莉高天文
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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