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Placental hematopoietic stem cell and preparation method thereof and placental hematopoietic stem cell injection

A technology for hematopoietic stem cells and placenta, applied in the field of placental hematopoietic stem cells and their preparation, and injection of placental hematopoietic stem cells, can solve the problems of affecting vitality, cell damage, rough grinding method, etc., and achieve the effect of increasing quantity and vitality

Active Publication Date: 2012-09-12
BOYALIFE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the grinding method is relatively rough, the cells cannot be completely separated from the tissue, and the number is small, and the grinding method will damage the cells and affect the vitality
The enzymatic method mostly uses a single enzyme to carry out enzymatic preparation, most of which are type IV collagenase, such as the Chinese patent application number 01131190.8 "A new method for extracting hematopoietic stem cells from placental tissue for establishing a hematopoietic stem cell bank" , which describes a protocol for preparing placental hematopoietic stem cells using a single collagenase, but does not disclose the number and viability of hematopoietic stem cells

Method used

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  • Placental hematopoietic stem cell and preparation method thereof and placental hematopoietic stem cell injection

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Experimental program
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Effect test

Embodiment 1

[0033] Embodiment 1: Preparation method of the present invention prepares placental hematopoietic stem cells

[0034] 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 storage bottle was prepared before the operation as a transport bottle, which contained commercially available cell culture medium, and was sent to the preparation place within 12 hours at 4°C after collection to separate and prepare placental hematopoietic stem cells.

[0035]In a sterile ultra-clean bench, take 1-2ml of placenta collection liquid for sterility testing, then remove the placental capsule, cut the placental tissue, put the chopped placental tissue into a beaker and wash it repeatedly with normal saline for 1-3 times , wash away the blood remaining on the surface of ...

Embodiment 2

[0043] Embodiment 2: the contrast of cell quantity and vitality

[0044] The placenta with the same source and volume as in Example 1 was used to prepare placental hematopoietic stem cells by using the existing grinding method and single collagenase method, and the steps involving resuspension, washing, centrifugation, etc. were all consistent with those in Example 1. In the single collagenase method, the amount and the mass percentage of collagenase of the single collagenase digestion solution adopted are all the same as the amount and the collagenase mass percentage of the corresponding collagenase digestion solution in Example 1, and finally these several methods are prepared The number and viability of the placental hematopoietic stem cells were detected, and the results are shown in Table 1.

[0045] Table 1 Cell number and viability

[0046]

[0047] It can be seen from the above table that the number of placental hematopoietic stem cells prepared by multiple collage...

Embodiment 3

[0048] Embodiment 3: Preparation method of the present invention prepares placental hematopoietic stem cells

[0049] 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 storage bottle was prepared before the operation as a transport bottle, which contained commercially available cell culture medium, and was sent to the preparation place within 12 hours at 4°C after collection to separate and prepare placental hematopoietic stem cells.

[0050] In a sterile ultra-clean bench, take 1-2ml of placenta collection liquid for sterility testing, then remove the placental capsule, cut the placental tissue, put the chopped placental tissue into a beaker and wash it repeatedly with normal saline for 1-3 times , wash away the blood remaining on the surface of...

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Abstract

The invention relates to the field of biology and discloses a placental hematopoietic stem cell and a preparation method thereof and a placental hematopoietic stem cell injection. The preparation method comprises the following steps of: after pretreating a placenta, digesting the treated placenta by using collagenase type IV digestive juice and collagenase type II digestive juice, then washing and filtering and respectively collecting first filtrate and first residue; digesting the first filter residue by using collagenase type I digestive juice and the collagenase type II digestive juice in the first step and then washing, filtering and collecting second filtrate; combining the filtrate from two times, centrifuging and discarding supernate, re-suspending and precipitating; adding a resuspension solution into a lymphocytes separation medium; and carrying out density gradient centrifugation, collecting a buffy coat cell solution on an intermediate layer and centrifuging and discarding supernate to obtain the placental hematopoietic stem cell. According to the placental hematopoietic stem cell and the preparation method thereof disclosed by the invention, collagenase type IV and collagenase type II are combined, collagenase type I and collagenase type II are combined and the hematopoietic stem cell is prepared by fully digesting the placenta according to the characteristics of different collagenases, so that the quantity and the vitality of the prepared hematopoietic stem cell are improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a placental hematopoietic stem cell, a preparation method thereof and a placental hematopoietic stem cell injection. Background technique [0002] Hematopoietic stem cells refer to a class of cells with self-renewal and multidirectional 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] Hematopoietic stem cell transplantation is currently widely used in malignant blood diseases (such as acute leukemia, chronic myelogenous leukemia, etc.), non-malignant refractory blood diseases (such as aplastic anemia, myelodysplastic syndrome, etc....

Claims

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Application Information

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IPC IPC(8): C12N5/0735A61K35/50A61K9/08A61P35/02A61P7/06A61P7/00A61P37/04A61P35/00
Inventor 李春波吕康涛
Owner BOYALIFE
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