Preparation method of placenta hematopoietic stem cells

A technology of hematopoietic stem cells and placenta, which is applied in the field of preparation of placental hematopoietic stem cells, can solve the problems of low activity rate of mononuclear cells, short time consumption, unsatisfactory cell number and vitality, etc., to increase the number of mononuclear cells and their living cells The effect of increasing the number, increasing the number and viability, and increasing the number of CD34 positive cells

Active Publication Date: 2015-06-17
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the separation and extraction methods of placental HSC mainly adopt mechanical method and mechanical enzymatic hydrolysis method, etc., but the extraction process of these two methods is complicated, time-consuming and costly, and is not suitable for large-scale production and preparation.
[0005] Chinese patent CN103789262A discloses a preparation method of clinically applied placental hematopoietic stem cells, which uses enzymatic hydrolysis perfusion to prepare hematopoietic stem cells, and the number of CD34 positive cells finally obtained is 1.2-1.6×10 9 , however, under the premise that the enzymatic hydrolysis time is extremely short, the effect data is basically impossi

Method used

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  • Preparation method of placenta hematopoietic stem cells

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Embodiment 1

[0038] Embodiment 1: the preparation method of placental hematopoietic stem cell described in the present invention

[0039] Placenta with negative test results for infectious diseases such as hepatitis, syphilis, and AIDS and without obstetrical complications were selected, and the informed consent of the puerpera was obtained and signed an informed consent form before the operation. A sterile polystyrene placenta collection box was prepared before the operation as a shipping container, which contained placental preservation solution, and was sent to the preparation place within 12 hours at 4°C after collection to separate and prepare placental hematopoietic stem cells.

[0040] In a sterile ultra-clean bench, take 1-2ml of placenta collection fluid for sterility testing, then remove the placenta from the amniotic membrane, wash the placenta with perfusate at 0-4°C, and use a disposable syringe to draw perfusate and infuse it into the placenta The two arteries until the serum...

Embodiment 2

[0044] Embodiment 2: the preparation method of placental hematopoietic stem cell described in the present invention

[0045] Placenta with negative test results for infectious diseases such as hepatitis, syphilis, and AIDS and without obstetrical complications were selected, and the informed consent of the puerpera was obtained and signed an informed consent form before the operation. A sterile polystyrene placenta collection box was prepared before the operation as a shipping container, which contained placental preservation solution, and was sent to the preparation place within 12 hours at 4°C after collection to separate and prepare placental hematopoietic stem cells.

[0046] In a sterile ultra-clean bench, take 1-2ml of placenta collection fluid for sterility testing, then remove the placenta from the amniotic membrane, wash the placenta with perfusate at 0-4°C, and use a disposable syringe to draw perfusate and infuse it into the placenta The two arteries until the serum...

Embodiment 3

[0050] Embodiment 3: the preparation method of placental hematopoietic stem cell described in the present invention

[0051] Placenta with negative test results for infectious diseases such as hepatitis, syphilis, and AIDS and without obstetrical complications were selected, and the informed consent of the puerpera was obtained and signed an informed consent form before the operation. A sterile polystyrene placenta collection box was prepared before the operation as a shipping container, which contained placental preservation solution, and was sent to the preparation place within 12 hours at 4°C after collection to separate and prepare placental hematopoietic stem cells.

[0052] In a sterile ultra-clean bench, take 1-2ml of placenta collection fluid for sterility testing, then remove the placenta from the amniotic membrane, wash the placenta with perfusate at 0-4°C, and use a disposable syringe to draw perfusate and infuse it into the placenta The two arteries until the serum...

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Abstract

The invention relates to the technical field of biology, and discloses a preparation method of placenta hematopoietic stem cells. The method comprises the following steps: cleaning a pretreated placenta by injection with an irrigating solution; injecting an enzymolysis solution from the placenta artery to the placenta vein until the enzymolysis solution flows out, tying the placenta artery and vein, and standing; loosening the placenta artery and vein, injecting the irrigating solution from the artery, collecting all the liquid at the vein, centrifugating, taking the cell precipitate, recovering and purifying single karyocytes comprising hematopoietic stem cells sequentially by a hydroxyethyl starch process and an erythrocyte cracking process, and resuspending to obtain the placenta hematopoietic stem cells. The method improves the enzymolysis injection technique, selects multiple mutually-suitable components to constitute the novel enzymolysis solution which is used in the placenta hematopoietic stem cell preparation process, can mobilize the hematopoietic stem cells inside the placenta to quickly enter the blood circulation system within a short time, and enhances the single karyocyte quantity comprising hematopoietic stem cells, the living cell quantity and the CD34 positive cell quantity.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preparing placental hematopoietic stem cells. Background technique [0002] Hemopoietic Stem cells (HSCs) are a group of primitive hematopoietic cells that exist in hematopoietic tissues. They are not tissue-fixed cells and can exist in hematopoietic tissues and blood. Hematopoietic stem cells are the origin of all hematopoietic and immune cells, with self-renewal and multilineage differentiation capabilities. Its basic feature is self-renewal ability, that is, after a cell cycle activity, it can produce two hematopoietic stem cells with the same properties as before division, and it also has multi-directional differentiation ability, that is, under certain environmental conditions, hematopoietic stem cells have The ability to differentiate into various blood cell lines. [0003] With the in-depth research on hematopoietic stem cells (HSC), HSC transplantation has beco...

Claims

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Application Information

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IPC IPC(8): C12N5/0789
Inventor 葛啸虎陈海佳王一飞卢瑞珊戴国胜马岩岩
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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