Human skin digestion complex enzyme and method for isolating micro-immune cell Treg cells from human few full-thickness skin

Abstract

The invention belongs to the field of medicine, and relates to human skin digestion complex enzyme and a method for isolating micro-immune cell Treg cells from human few full-thickness skin; the human skin digestion complex enzyme is 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 final concentration of 1%, type 4 collagenase with the final concentration of 4 mg / ml, hyaluronidase with the final concentration of 0.5 mg / ml, and deoxyribonuclease with the final concentration of 2 mg / ml; the method for isolating micro-immune cell Treg cells from human few full-thickness skin includes the following steps: true epidermal separation; digestion with a human skin digestion complex enzyme solution; performing filtration; and performing analytical analysis using flow cytometry. The invention facilitates the research of cytological and molecular studies on skin-specific Treg cells, and can also provide some reference for isolating 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 human skin digestive complex enzyme and a method for isolating trace immune cells Treg cells from a small amount of full-thickness human skin. Background technique [0002] As the first protective barrier of the human body, the skin can perform a series of functions to protect us, such as preventing the invasion of pathogens and rapidly healing the skin. All these processes need to be completed with the assistance of the immune system, so the skin is complex and Huge library of immune cells. 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, so study and elucidate the molecular and cellular aspects of different skin diseases Mechanisms are an essential avenu...

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Problems solved by technology

[0004] In 2004, Patrick reported the method of isolating T cells from human skin tissue. After the tissue was shredded mechanically, it was digested with enzymes, then filtered, and T cells were obtained by Ficoll-Paque gradient centrifugation. However, the separation of this method The principle is mainly based on the principle of enzymatic digestion and the properties of different cells. Its realization requires high enzyme digestibility and a large cell base. The accuracy is controversial, and it cannot be applied to the isolation of extremely low proportions like Treg cells. cell population

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