In vitro isolation and amplification method and application of human peripheral blood Treg cells

Abstract

The invention relates to an in vitro isolation and amplification method and an application of human peripheral blood Treg cells, and belongs to the technical fields of in vitro isolation and amplification methods and applications of cells. The method comprises the following steps: 1) collecting peripheral blood of the mate of an unexplained recurrent spontaneous abortion patient, and separating peripheral blood mononuclear cells by gradient centrifugation; 2) washing the peripheral blood mononuclear cells twice with a phosphate buffer solution, and counting the obtained mononuclear cell; 3) separating CD4<+>T cells and Treg cells by a magnetic bead sorting technology; and 4) amplifying the Treg cells. The method allows the Treg cells with a high purity to be obtained, the number of cells,required for clinical application, is obtained by the in vitro amplification method, and the human peripheral blood Treg cells obtained in the invention can be used for treating the unexplained recurrent spontaneous abortion, and can significantly improve the pregnant rate of unexplained recurrent miscarriage patients.

Description

technical field [0001] The invention relates to an in vitro separation and expansion method of human peripheral blood Treg cells and its application, and belongs to the technical field of the in vitro separation and expansion method of cells and its application. Background technique [0002] Recurrent Spontaneous Abortion (RSA) refers to two or more consecutive spontaneous abortions with the same sexual partner. The incidence rate is about 5% in women of childbearing age, and the rate of re-pregnancy failure in such patients is as high as 50%. The etiology of RSA is complex, clinically about 50% are related to chromosomal abnormalities, reproductive system anatomical abnormalities, endocrine abnormalities, infection and thrombosis tendency, and another 50% of RSA patients have unknown etiology, which is called unexplained recurrent abortion (URSA), which is considered to be related to It is related to maternal-fetal immune rejection and is a refractory infertility that needs...

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Problems solved by technology

6) The level of Treg cells, the ability to multiply in vitro and the secretion of cytokines during the implantation period are closely related to the success or failure of the implantation, and the expression of Treg cells in the peripheral blood of patients with repeated implantation failures decreased during the implantation period (Wang et al., 2017)

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