Method for improving directional differentiation effects on erythroid progenitor cells in umbilical cord blood

An erythroid progenitor cell and directional differentiation technology is applied in the field of improving the directional differentiation effect of umbilical cord blood erythroid progenitor cells, which can solve problems such as increasing costs, restricting clinical use, etc. Effects of toxic effects

Inactive Publication Date: 2016-07-13
广州市天河诺亚生物工程有限公司
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If a bioreactor is used, the production of erythroid progenitor cells can indeed be greatly increased, but the subsequent substantial increase in cost also restricts clinical use

Method used

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  • Method for improving directional differentiation effects on erythroid progenitor cells in umbilical cord blood
  • Method for improving directional differentiation effects on erythroid progenitor cells in umbilical cord blood
  • Method for improving directional differentiation effects on erythroid progenitor cells in umbilical cord blood

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Experimental program
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Effect test

Embodiment 1

[0050] Umbilical cord blood was collected from healthy pregnant women and infants, and the tests of 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:

[0051] 1. Preparation of Piceatannol solution and basic expansion medium

[0052] Piceatanol (≥99%, GC), dissolved in a cell protection agent, prepared into a concentration of 10mmol / L Piceatanol storage solution;

[0053] Basic expansion medium: Stemspan medium supplemented with cytokines IL-3 (final concentration 20ng / mL), EPO (final concentration 1.5U / mL), SCF (final concentration 100ng / mL) and DEX (final concentration is 1nM);

[0054] Expansion medium containing picetanol: adding the above-prepared picetanol storage solution to the basic expansion medium to obt...

Embodiment 2

[0089] Directed expansion of umbilical cord blood hematopoietic stem cells to erythroid progenitor cells according to the method of Example 1:

[0090] After 21 days of culture, the final result: the number of hematopoietic stem / progenitor cells in Control and 5 μmol / L, 10 μmol / L, 15 μmol / L piceatanol samples were: 39.02×10 6 pcs, 44.29×10 6 pcs, 40.25×10 6 pcs, 47.52×10 6 compared with the first day, the cell number ratios were: 24.85 times, 28.21 times, 25.63 times, 30.26 times.

[0091] Detected by flow cytometry, in CD71 + In terms of cell ratio, the results of each sample are: 73.97%, 67.18%, 66.65%, 68.6%;

[0092] CD71 + In terms of the total number of cells, the results of the samples with different concentrations of piceatanol were: 28.86×10 6 pcs, 29.76×10 6 pcs, 26.83×10 6 pcs, 32.6×10 6 compared with the first day, the cell number ratios were: 42.07 times, 43.37 times, 39.1 times, 47.51 times.

[0093] In terms of the number of BFU-E colonies, the results...

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Abstract

The invention belongs to the technical field of cells, and particularly relates to a method for improving directional differentiation effects on erythroid progenitor cells in umbilical cord blood.The method includes directionally differentiating and amplifying specimen cells by the aid of amplification cultivation media with piceatannol which is a natural compound.Hematopoietic stem cells in the umbilical cord blood are used as the specimen cells.The method has the advantages that directional differentiation operation on the erythroid progenitor cells can be easily, conveniently, safely and efficiently carried out by the aid of the amplification cultivation media with the piceatannol; the piceatannol and cytokines are simultaneously added into the hematopoietic stem cells in procedures for directionally differentiating and cultivating the hematopoietic stem cells to obtain the erythroid progenitor cells, accordingly, effects of increasing the quantities of the generated erythroid progenitor cells and improving BFU-E (burst forming unit-erythroid) colony formation ability can be realized, the method is easy and convenient to implement and low in cost, and the progenitor cells are high in safety and red blood cell differentiation capacity.

Description

technical field [0001] The invention belongs to the technical field of cells, and in particular relates to a method for improving the directional differentiation effect of umbilical cord blood erythroid progenitor cells. Background technique [0002] In the course of clinical treatment, blood transfusion is an important therapeutic aid, and red blood cells are the most commonly used blood transfusion components. The domestic clinical blood supply relies on voluntary donations, but while the demand continues to expand, the growth of voluntary blood donations is relatively backward. This has caused blood shortages in many cities across the country, and there are gaps in the supply of both whole blood and red blood cells and other blood components. [0003] In order to make up for the insufficient supply of red blood cells caused by the relatively lagging increase in voluntary blood donations, researchers have conducted research on red blood cell substitutes. The main red bloo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078C12N5/0789
CPCC12N5/0641C12N2500/30C12N2506/11
Inventor 魏伟嵐山芮袁嘉恩
Owner 广州市天河诺亚生物工程有限公司
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