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Culture medium and method for promoting induced differentiation of hematopoietic stem cells into megakaryocyte by shake culture

A technology of hematopoietic stem cells and inducing differentiation, applied in the field of bioengineering, can solve the problems of low efficiency of inducing differentiation and slow proliferation, etc.

Pending Publication Date: 2021-06-29
ACADEMY OF MILITARY MEDICAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the current system for promoting the induction and differentiation of hematopoietic stem cells into megakaryocytes still has problems such as slow proliferation and low induction differentiation efficiency.

Method used

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  • Culture medium and method for promoting induced differentiation of hematopoietic stem cells into megakaryocyte by shake culture
  • Culture medium and method for promoting induced differentiation of hematopoietic stem cells into megakaryocyte by shake culture
  • Culture medium and method for promoting induced differentiation of hematopoietic stem cells into megakaryocyte by shake culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] 1. Materials

[0052] Cord blood mononuclear cells: mix cord blood with 6% hydroxyethyl starch, settle red blood cells for 20-30 minutes, take supernatant, centrifuge, collect cell pellets, resuspend cell pellets in normal saline, add human lymphocyte separation medium , centrifuged, and the cell pellet was collected to obtain umbilical cord blood mononuclear cells.

[0053] st36pSR1 medium: containing 50ng / mL thrombopoietin, 50ng / mL recombinant human stem cell factor, 20ng / mL interleukin-3, 50ng / mL interleukin-6, 1mg / mL poloxamer 188, 1μM Aryl hydrocarbon receptor antagonist stemspan serum-free medium.

[0054] St36 medium: stemspan serum-free medium containing 50ng / mL of thrombopoietin, 50ng / mL of recombinant human stem cell factor, 20ng / mL of interleukin-3, and 50ng / mL of interleukin-6.

[0055] st36SR1 medium: stemspan containing 50 ng / mL thrombopoietin, 50 ng / mL recombinant human stem cell factor, 20 ng / mL interleukin-3, 50 ng / mL interleukin-6, 1 μM aryl hydrocar...

Embodiment 2

[0067] 1. Materials

[0068] Umbilical cord blood mononuclear cells and st36pSR1 medium are the same as in Example 1, and will not be repeated here.

[0069] 2. Experimental method

[0070] Cord blood mononuclear cells were divided into 4 groups on average, namely st36pSR1 static group, st36pSR1 shaking group, st36pSR1+GM-CSF+IL-11 static group, st36pSR1+GM-CSF+IL-11 shaking group. in:

[0071] The static group of st36pSR1 adopts G-Rex culture device to press 2×10 5 piece / cm 2 Inoculated in st36pSR1 medium, at 37°C, 5% by volume CO 2 Culture in the incubator statically, change half of the medium on 7d, 12d, and 17d, supplement with thrombopoietin, recombinant human stem cell factor, interleukin-3, interleukin-6, poloxamer 188 and aromatic hydrocarbon receptor antagonist stemspan culture medium, so that the culture medium after changing the medium contains: 50ng / mL of thrombopoietin, 50ng / mL of recombinant human stem cell factor, 20ng / mL of interleukin-3, 50ng / mL of interl...

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Abstract

The invention provides a culture medium and a method for promoting induced differentiation of hematopoietic stem cells into megakaryocyte cells. The culture medium comprises a basic culture medium; a thrombopoietin; recombinant human stem cell factors; Interleukin-3; Interleukin-6; a poloxamer 188; and an aromatic hydrocarbon receptor antagonist. The culture medium of the invention can effectively promote induced differentiation of hematopoietic stem cells to megakaryocyte cells, improves the differentiation efficiency and the yield and activity of megakaryocyte cells, and has wide application prospects.

Description

technical field [0001] The invention relates to the field of bioengineering. Specifically, the present invention relates to a culture medium and a method for shaking hematopoietic stem cells to induce differentiation into megakaryocytes. Background technique [0002] Platelets are formed components in blood, have the functions of adhesion, aggregation and release, participate in hemostasis, maintain the integrity of blood vessel walls, coagulation and inflammatory reactions, and play an important role in the development of diseases. Thrombocytopenia can be caused by a variety of reasons, such as tumors, aplastic anemia, thrombocytopenic purpura, etc., and platelet transfusion is often used as an adjuvant supportive treatment. Hematopoietic stem / progenitor cells have the potential of self-renewal and multi-lineage differentiation. Simulating or partially simulating the hematopoietic process in vivo can make them directional induce differentiation into megakaryotic cells, whi...

Claims

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Application Information

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IPC IPC(8): C12N5/078C12N5/0789
CPCC12N5/0644C12N5/0647C12N2501/145C12N2501/125C12N2501/2303C12N2501/2306C12N2500/50C12N2501/60Y02A50/30
Inventor 裴雪涛岳文陈琳李艳华习佳飞姚海雷刘晓丹李雪吕洋范增何丽娟谢小燕张博文南雪林玉环
Owner ACADEMY OF MILITARY MEDICAL SCI
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