Preparation method of post-resuscitation protective agent for refrigerated umbilical cord blood hematopoietic stem cells

A technology of hematopoietic stem cells and preparation methods, which is applied in the field of preparation of protective agents after resuscitation of umbilical cord blood hematopoietic stem cells, which can solve the problems of increased cell fragments, reduced number of cells, easy aggregation of cells, etc., achieving less dosage and convenient purchase and use , the effect of protective activity

Active Publication Date: 2019-06-21
天晴干细胞股份有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to solve the problems that the umbilical cord blood hematopoietic stem cells are easily agglomerated, the cell fragments increase, the viability and the number of cells are reduced in the pre-treatment of the umbilical cord blood hematopoietic stem cells that are resuscitated and frozen with physiological saline, and a method for freezing is provided. Preparation method of protective agent after resuscitation of umbilical cord blood hematopoietic stem cells

Method used

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  • Preparation method of post-resuscitation protective agent for refrigerated umbilical cord blood hematopoietic stem cells
  • Preparation method of post-resuscitation protective agent for refrigerated umbilical cord blood hematopoietic stem cells
  • Preparation method of post-resuscitation protective agent for refrigerated umbilical cord blood hematopoietic stem cells

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specific Embodiment approach 1

[0013] Specific embodiment 1: In this embodiment, a preparation method for protecting agent after resuscitation of frozen umbilical cord blood hematopoietic stem cells is carried out according to the following steps: 1. Configure buffer: weigh 3-3.5 g of sodium chloride, potassium chloride 0.1~0.2g, magnesium chloride 0.1~0.2g, sodium acetate 1~1.5g, sodium citrate 0.2~0.3g, calcium gluconate 0.3~0.4g and trehalose 0.2~0.3g; then sodium chloride, chloride Potassium, magnesium chloride, and sodium citrate were dissolved in 480-520mL of deionized water in turn, stirred until completely melted, added calcium gluconate and trehalose, stirred until dissolved, stood still for 8-12 minutes, added sodium acetate, stirred evenly, stood still for 4- 6min, adjust the pH value between 7.0-7.2 to obtain a buffer solution; 2. Fibrin-depleted plasma: add calcium chloride solution to the plasma, shake for 4-6min, centrifuge through a density gradient, take the liquid phase, and obtain the defi...

specific Embodiment approach 2

[0015] Specific embodiment two: the difference between this embodiment and specific embodiment one is: in step one, take by weighing 3.12g of sodium chloride, 0.15g of potassium chloride, 0.11g of magnesium chloride, 1.025g of sodium acetate, 0.254g of sodium citrate, Calcium gluconate 0.322g and trehalose 0.25g. Others are the same as the first embodiment.

specific Embodiment approach 3

[0016] Specific embodiment 3: This embodiment differs from specific embodiment 1 or 2 in that: in step 1, sodium chloride, potassium chloride, magnesium chloride, and sodium citrate are dissolved in 500 mL of deionized water in sequence. Others are the same as those in Embodiment 1 or 2.

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Abstract

The invention discloses a preparation method of a post-resuscitation protective agent for refrigerated umbilical cord blood hematopoietic stem cells, and aims to solve problems that in existing pre-treatment of resuscitating the refrigerated umbilical cord blood hematopoietic stem cells by using normal saline, the cells are easy to be agglomerated, cell debris is increased, a survival rate is reduced and the number of the cells is reduced. The method comprises the following steps: one, preparing buffer solution; two, preparing plasma without fibrin; and three, adding low-molecular-weight heparin sodium, hydroxyethyl starch and polyethylene glycol to the buffer solution; and finally adding the plasma without fibrin, completing. The protective agent is capable of effectively protecting the survival rate of the cells, wherein the cells are not easy to be agglomerated, and cell colony formation is better, effectively protecting activity of the umbilical cord blood hematopoietic stem cellsafter resuscitation, and the number of the cells, wherein the cells are not agglomerated, and guaranteeing a formation force of a colony. The protective agent is applied to the preparation field of the post-resuscitation protective agent for the hematopoietic stem cells.

Description

technical field [0001] The invention relates to a preparation method of a protective agent for frozen umbilical cord blood hematopoietic stem cells after resuscitation. Background technique [0002] Umbilical cord blood refers to the placenta and the blood in blood vessels on the side of the umbilical cord close to the fetus obtained by collecting and draining the placenta and umbilical cord when the umbilical cord is ligated and cut off after the newborn is cut off or punctured. Cord blood mesenchymal stem cells and hematopoietic stem cells are abundant. Umbilical cord blood hematopoietic stem cells are primitive cells with strong differentiation potential and the ability to form colonies and proliferate and differentiate. Induced or affected by specific factors, cord blood hematopoietic stem cells can differentiate into various tissues or cells. CD34 + The proportion of nucleated cells in umbilical cord blood is about 1%, which is similar to bone marrow, and its content ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0789
Inventor 王灵娟张怡刘艳青
Owner 天晴干细胞股份有限公司
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