Induction method for improving capacity of mesenchymal stem cells in promoting acute myeloid leukemia cell differentiation
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A mesenchymal stem cell and cell technology, applied in the field of induction to improve the ability of mesenchymal stem cells to promote the differentiation of acute myeloid leukemia cells, can solve the problems that clinical applications have not been well studied
Active Publication Date: 2020-07-28
NANJING UNIV
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Despite this, the role of BMP6 in the treatment of human dis...
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Embodiment 1
[0026] Isolation and subculture of umbilical cord mesenchymal stem cells
[0027] 1. Prepare tissue digestion enzymes (type II collagenase 250U / ml, dispase 100U / ml, hyaluronidase 10U / ml), dissolve in α-MEM medium at 37°C, filter and sterilize with a 0.22μm filter for later use ;
[0028] 2. Take the umbilical cord tissue of the newborn baby, put the umbilical cord into a petri dish, wash it with PBS containing 0.1% penicillin-streptomycin double antibody, put it into α-MEM medium, peel off the three blood vessels, and cut the umbilical cord into pieces 1-2mm 3 organization block;
[0029] 3. Mix the shredded tissue pieces with the prepared tissue digestion enzyme solution in a volume ratio of 1:1 in a 50ml centrifuge tube, digest at 37°C, 200rpm for 3 hours, and wait until the tissue pieces are basically completely digested;
[0030] 4. Centrifuge the digested tissue fluid at 300 g for 5 min at 4°C and discard the supernatant. PBS was resuspended in 50ml α-MEM medium, cent...
Embodiment 2
[0034] Optimization of UC-MSCs activation protocol
[0035]1. BMP6 was dissolved in PBS (pH=7.4) at a concentration of 5 μM.
[0036] 2. Add 5 μM biological BMP6 into the complete medium at a ratio of 1:1000 to a final concentration of 5 nM.
[0037] 3. After UC-MSCs grow to 70-80% abundance, add medium containing 5nM BMP6 and culture for 24 hours.
[0038] 4. After activating UC-MSCs for 24 hours, discard the supernatant, wash twice with PBS of pH 7.4, and continue culturing with α-MEM medium containing 10% FBS.
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Abstract
The invention belongs to the technical field of medical biology, and particularly relates to an application of an induction method for improving the capacity of mesenchymal stem cells in promoting acute myeloid leukemia cell differentiation. The mesenchymal stem cells provided by the invention are human-derived umbilical cord mesenchymal stem cells (UC-MSCs), and mesenchymal stem cells (pre-activeMSCs) obtained after pretreatment and activation by cytokine bone morphogenetic protein 6 (BMP6); the pre-active MSCs can efficiently induce the acute myeloid leukemia cell differentiation, and the capacity is achieved by changing the expression quantity of IL-6 and IDO of the UC-MSCs; and the mesenchymal stem cells are allogeneic mesenchymal stem cells, the secretion capacity of the mesenchymalstem cells can be remarkably changed after the mesenchymal stem cells are treated and activated by the BMP6, and the AML resistance of the mesenchymal stem cells is remarkably enhanced. Compared withchemotherapy, mesenchymal stem cell transplantation has lower toxic and side effects, has low immunogenicity and meets clinical requirements. According to the method, the capacity of the MSCs in inducing the acute myelogenous leukemia cell differentiation is improved by adopting the method of pretreating and activating the MSCs through the BMP6, and the method can be applied to differentiation treatment of acute myelogenous leukemia.
Description
technical field [0001] The invention relates to the field of medical and biological technologies, in particular to an induction method for improving the ability of mesenchymal stem cells to promote the differentiation of acute myeloid leukemia cells. Background technique [0002] Mesenchymal stem cells (mesenchymal stem cells, MSCs) were first discovered in the bone marrow, and later found that mesenchymal stem cells exist in almost all tissues during the growth and development of the human body. Currently, mesenchymal stem cells can be isolated and prepared from tissues such as bone marrow, adipose tissue, umbilical cord, placenta and amniotic fluid. MSCs not only have strong self-renewal ability and multi-directional differentiation potential, but also have the characteristics of immune regulation ability and low immunogenicity. They are regeneration source cells with broad application prospects in the field of transplantation. Favored by scientists and medical researcher...
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