A method for increasing the number of directional differentiation of umbilical cord blood megakaryotic progenitor cells

A technology for megakaryotic progenitor cells and directed differentiation, which is applied in the field of increasing the number of directed differentiation of umbilical cord blood hematopoietic stem cells to megakaryotic progenitor cells, can solve problems such as increased cost, pollution, and adverse effects on in vitro amplification efficiency, and achieves the goal of increasing the number of directed differentiation, The effect of high safety and less toxic effects

Active Publication Date: 2017-10-13
广州市天河诺亚生物工程有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Complex composition adversely affects in vitro amplification efficiency
Although purer hematopoietic stem cells can be obtained by magnetic bead sorting to optimize in vitro expansion, the increased cost greatly restricts the situation that requires a large amount of expansion, and more operating steps also bring greater risk of contamination

Method used

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  • A method for increasing the number of directional differentiation of umbilical cord blood megakaryotic progenitor cells
  • A method for increasing the number of directional differentiation of umbilical cord blood megakaryotic progenitor cells
  • A method for increasing the number of directional differentiation of umbilical cord blood megakaryotic progenitor cells

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Umbilical cord blood was collected from healthy pregnant women and infants. Hepatitis B, hepatitis C, syphilis, AIDS, cytomegalovirus, TORCH, mycoplasma, chlamydia, G-6PD and thalassemia were all negative. The transport conditions of the specimens from collection to return to the blood bank were kept at 4-8°C. Perform directed differentiation of megakaryotic progenitor cells as follows:

[0034] 1. Preparation of resveratrol solution and universal medium

[0035] Resveratrol (≥99%, GC) was dissolved in a cell protection agent, and prepared into a resveratrol storage solution with a concentration of 10mmol / L.

[0036] The cell protection agent is a mixture obtained by mixing DMSO and Dextron at a volume ratio of 1:1.

[0037] Specific medium: Stemspan medium from STEMCELL company, IL-3 (20ng / mL), IL-6 (50ng / mL), SCF (50ng / mL) and TPO (50ng / mL) cytokines were added respectively.

[0038]Specific medium containing resveratrol: Add the above-prepared resveratrol storage...

Embodiment 2

[0059] Directed expansion of umbilical cord blood hematopoietic stem cells to megakaryotic progenitor cells with reference to the method of Example 1:

[0060] On the first day, the initial number of hematopoietic stem / progenitor cells in Control, 5 μmol / L, 15 μmol / L, and 25 μmol / L resveratrol samples were: 4.26×10 6 pcs / mL, 4.0×10 6 pcs / mL, 3.54×10 6 pcs / mL, 3.88×10 6 individual / mL.

[0061] On the 6th day, the number of hematopoietic stem / progenitor cells in Control, 5 μmol / L, 15 μmol / L, and 25 μmol / L resveratrol samples were: 3.34×10 6 pcs / mL, 3.73×10 6 pcs / mL, 4.27×10 6 pcs / mL, 4.46×10 6 cells / mL; detected by flow cytometry, in CD41 + In terms of cell ratios, the results were: 6.35%, 8.18%, 8.45%, and 8.54%, respectively.

[0062] On the 12th day, the number of hematopoietic stem / progenitor cells in Control, 5 μmol / L, 15 μmol / L, and 25 μmol / L resveratrol samples were: 15.57×10 6 pcs / mL, 16.36×10 6 pcs / mL, 17.64×10 6 pcs / mL, 11.39×10 6 cells / mL, compared with th...

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Abstract

The invention discloses a method for increasing the number of directional differentiation of umbilical cord blood megakaryocyte progenitor cells. In this method, umbilical cord blood hematopoietic stem cells are used as sample cells, and directed differentiation is carried out in a specific medium containing resveratrol. The specific medium containing resveratrol makes the operation of directed differentiation of megakaryotic progenitor cells easier, safer and more efficient. In the process of directional differentiation and culture of umbilical cord blood hematopoietic stem cells to megakaryotic progenitor cells, resveratrol is added together with cytokines to achieve the effect of increasing the number of megakaryotic progenitor cells. The method of the present invention is easy to operate and low in cost. Cell safety is high.

Description

technical field [0001] The invention belongs to the field of cell technology, and in particular relates to a method for increasing the number of directed differentiation of umbilical cord blood hematopoietic stem cells to megakaryotic progenitor cells. Background technique [0002] Hematopoietic stem cells have the ability to self-renew and differentiate into precursor cells of various blood cells, which eventually form blood cells, including red blood cells, white blood cells and platelets. Hematopoietic stem cells are mainly derived from bone marrow, umbilical cord blood and peripheral blood, and are clinically used for transplantation to treat various malignant blood diseases and certain immune diseases. [0003] In 1989, the first case of umbilical cord blood transplantation was successful. So far, umbilical cord blood transplantation / transfusion has become an important clinical treatment for blood system diseases and certain immune diseases. Like bone marrow hematopoie...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0789
Inventor 嵐山芮魏伟袁嘉恩
Owner 广州市天河诺亚生物工程有限公司
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