Preparing method for clinic level cells for enhancing immune modulating function of MSCs

An immune regulation and cell technology, applied in the direction of animal cells, vertebrate cells, bone/connective tissue cells, etc., can solve the problems of low yield and limited immune regulation ability, achieve immune regulation and promotion, save raw material costs, and increase proliferation ability and the ability to resist NK cell killing

Active Publication Date: 2015-12-16
丛秀丽
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Problems solved by technology

[0023] The technical problem to be solved by the present invention is to overcome the defects of low yield and limited immunoregulatory ability caused by the use of medium contai...

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  • Preparing method for clinic level cells for enhancing immune modulating function of MSCs
  • Preparing method for clinic level cells for enhancing immune modulating function of MSCs
  • Preparing method for clinic level cells for enhancing immune modulating function of MSCs

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Embodiment

[0054] A clinical level cell preparation method for enhancing the immune regulation function of MSCs, comprising the following steps:

[0055] 1. Umbilical cord collection and mesenchymal stem cell isolation and culture

[0056] (1) Take umbilical cords of 10-15 cm from normal full-term healthy newborns, and immediately put them into DMEM medium with 10-20% mass fraction of double antibody solution; the umbilical cords need to be processed within 8-12 hours;

[0057] (2) Take out the umbilical cord in the ultra-clean bench, disinfect the surface of the umbilical cord with 75% alcohol, and fully wash the residual blood with normal saline; cut the umbilical cord into 1-2 cm small pieces and rinse again; cut open the umbilical cord, and draw out the umbilical vein and umbilical artery , cut the remaining Wharton's jelly umbilical cord tissue into 1-2mm 3 Size, inoculated at the bottom of T75 culture flask with an interval of 0.5-1.0 cm, and incubated the culture flask in a CO2 i...

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Abstract

The invention discloses a preparing method for clinic level cells for enhancing the immune modulating function of MSCs. The preparing method is suitable for culturing the mesenchymal stem cells from bone marrow, fat and the umbilical cord. A serum-free complete medium is adopted to culture the mesenchymal stem cells from umbilical cord tissue, a traditional method is improved by applying a secondary inoculation method, and the obtaining rate of primary cells is increased. Imidazoquinoline R848 with the concentration of 1-5 micrograms/milliliters and recombinant human interferon I FN-alpha-2b with the concentration of 100-200 units/milliliters are applied as pretreatment factors and obviously induce high expression of the MSCs and secretory cell factors VEGF, HGF and PGE-2, the capacity for modulating immune and promoting tissue regeneration of the MSCs is brought into play better, the multiplication capacity and the capacity for resisting NK cell killing of the MSCs can be improved obviously, and the pretreated MSCs have the enhanced capacity for suppressing lymphocyte proliferation and can be applied to treatment on the GVHD with T cells as the principal and other autoimmune diseases better.

Description

technical field [0001] The invention relates to the application field of stem cells, in particular to a method for preparing clinical-grade cells for enhancing the immune regulation function of MSCs. Background technique [0002] Mesenchymal stem cells: Mesenchymal stem cells (mesenehymalstemcells, MSCs) are pluripotent stem cells derived from early mesoderm with high self-renewal and multidirectional differentiation potential. A variety of tissue cells, such as bone marrow stromal cells, osteoblasts, chondrocytes, adipocytes, myoblasts, and astrocytes. Recent studies have shown that, in addition to supporting hematopoiesis in vitro and promoting hematopoietic reconstruction in vivo, MSCs can also inhibit allogeneic immune responses and reduce graft-versus-host disease (GVHD). [0003] Human umbilical cord mesenchymal stem cells (hUCMSC): The human umbilical cord is connected between the mother and the fetus, and provides nutrition for the fetus during pregnancy. It consist...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 丛秀丽
Owner 丛秀丽
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