Method for efficiently and sequentially inducing macrophages to be polarized in vitro

Abstract

The invention discloses a method for efficiently and sequentially inducing macrophages to be polarized in vitro. The method comprises the following steps of: S1, carrying out multiplication culture onTHP-1 cells by using an RPMI1640 culture medium containing 10% of FBS and beta-mercaptoethanol; S2, carrying out pretreatment before induction on the THP-1 cells by using PBS containing HPL and a culture medium; S3, in an RPMI1640 culture medium which contains 5% of HPL and does not contain the beta-mercaptoethanol, slowly inducing the THP-1 cells by adopting low-concentration PMA so as to enablethe THP1 cells to be differentiated into M0 macrophages in an adherence mode, and then removing the PMA; and S4, in an RPMI1640 culture medium which contains 5% of HPL and does not contain the beta-mercaptoethanol, inducing M0 macrophages for more than 72 hours by using LPS/IFN-gamma, so that the M0 macrophages are polarized into M1; or inducing the M0 macrophages for more than 72 hours by usingIL-4/IL-13, so that the M0 macrophages are polarized into M2 macrophages. Compared with the prior art, the method disclosed by the invention has the advantages that firstly, the THP-1 cells are slowlyinduced by adopting the low-concentration PMA, so that the immatureness state of the M0 type macrophages differentiated from the THP1 cells can be guaranteed; and then the optimal induced polarization time of the subsequent M1/M2 type macrophages is over 72 hours, so that the polarization state of the M1/M2 type macrophages is guaranteed. Meanwhile, the differentiation process is carried out in an environment without animal-derived components, so that the polarization efficiency of the macrophages can be effectively ensured.

Description

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AI Technical Summary

Benefits of technology

This patented technology allows scientists to quickly stimulate large numbers of tiny red blood cells called Theonella monocytes through an enzyme process without affecting their ability to divide them correctly. By doing this they could help researchers better identify specific subpopulations of white blood cells like neutrocyte intermediating zone plasma cells. These isolated populations were then used to investigate how these special properties might change during inflammations caused by various diseases including autoimmune disorders.

Problems solved by technology

This patented technical problem addressed in this patents relates to improving the efficiency at producing large amounts of certain macromolecular substances called extracelluarbodies (Ebs). These EBs play key roles in both innate and adaptive immunocompetentiating processes involving inflammations, cancer development, autoimmune response, and regulation of various biological reactions like wound healing and tissue injury. Current techniques involve modifying these materials with chemical agents or enhancer compounds, but their effects on production remain uncertain due to lack of knowledge regarding how much each material contributes towards its own activity.

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