Preparation method for placenta hemopoietic stem cell

A technology of hematopoietic stem cells and placenta, applied in the field of preparation of placental hematopoietic stem cells, can solve the problems of unsatisfactory effect, small number of cells, low content of hematopoietic stem cells, etc., and achieve the effect of increasing the number of mononuclear cells, increasing the number and the activity rate

Active Publication Date: 2015-07-15
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The number of cells obtained by the mechanical method is small, and contains a large amount of cell debris, and the cell viability is low; while the mechanical enzymatic method can obtain a sufficient number of cells, but the content of hematopoietic stem cells is low
In addition, the entire extraction process is complicated, time-consuming, and costly; the extracted cells have low viability
[0005] U.S. Patent US 20120177616 "Methods for augmentation collection of placental hematopoietic stem cells and uses thereof" discloses a method for obtaining h

Method used

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  • Preparation method for placenta hemopoietic stem cell
  • Preparation method for placenta hemopoietic stem cell
  • Preparation method for placenta hemopoietic stem cell

Examples

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Effect test

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 biotechnical field and discloses a preparation method for a placenta hemopoietic stem cell. The preparation method comprises the following steps: irrigating and cleaning a pre-processed placenta with an irrigating solution; irrigating a mobilizing agent from placental arteries until the mobilizing agent stops flowing from the placental veins and the placental arteries and veins are ligatured and stood; loosening the placental arteries and veins and grouting the same amount of mobilizing agent from the arteries, collecting all liquids from the veins, and centrifuging to get cell sediments; then, recycling and purifying single karyocyte comprising the hemopoietic stem cell through a hydroxyethyl starch process and a red blood cell lysis process in sequence, and re-suspending to obtain the placenta hemopoietic stem cell. According to the preparation method, the mobilizing agent irrigating process is improved, a plurality of mutually suitable components are selected to form a novel mobilizing agent for being applied to a preparation process of the placenta hemopoietic stem cell, the placenta hemopoietic stem cell inside the placenta can be mobilized and can quickly enter a blood circulating system, and the quantity of single karyocyte comprising the hemopoietic stem cell as well as quantity of living cells of the single karyocyte and quantity of CD34 positive cells is increased.

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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