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Method for preparing dental pulp mesenchymal stem cells and establishing dental pulp mesenchymal stem cell bank and cell resuscitation method

A technology for stem cells and dental pulp, applied in the field of stem cell bank preparation, can solve the problems of small number of cells, high preparation cost, incomplete digestion of tissue blocks, etc., and achieve the effect of large cell loss and avoiding loss.

Inactive Publication Date: 2015-05-20
天晴干细胞股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The culture of cells by the tissue block method has a long culture cycle and a small number of cells; the number of cells cultured by the collagenase digestion method is small, the tissue block is not fully digested, and the preparation cost is high

Method used

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  • Method for preparing dental pulp mesenchymal stem cells and establishing dental pulp mesenchymal stem cell bank and cell resuscitation method
  • Method for preparing dental pulp mesenchymal stem cells and establishing dental pulp mesenchymal stem cell bank and cell resuscitation method
  • Method for preparing dental pulp mesenchymal stem cells and establishing dental pulp mesenchymal stem cell bank and cell resuscitation method

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specific Embodiment approach 1

[0054] Specific embodiment one: a kind of method for preparing dental pulp mesenchymal stem cells and establishing dental pulp mesenchymal stem cell bank of the present embodiment, it is carried out according to the following steps:

[0055] 1. Semi-digestion treatment of trypsin substitute:

[0056] 1) The pulp is taken out from the pulp cavity, and after aseptic treatment, the pulp is cut into small pieces;

[0057] 2) Put the above-mentioned cut dental pulp into a sterile EP tube, then add trypsin substitute into the EP tube, and then put the EP tube containing the tissue block and trypsin substitute into a 37°C incubator Medium digestion for 15-20 minutes, shaking the EP tube every 5 minutes;

[0058] 2. Primary culture of dental pulp tissue:

[0059] 3) adding the pulp tissue block digested in step 2 to the complete medium of mesenchymal stem cells, blowing and mixing thoroughly to obtain the pulp tissue block suspension;

[0060] 4) Inoculate the pulp tissue block sus...

specific Embodiment approach 2

[0074] Specific embodiment 2: The difference between this embodiment and specific embodiment 1 is that the pancreatic enzyme substitute is a recombinant enzyme, which does not contain animal-derived components, and can be stored at 4°C, and the components are stable. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0075] Embodiment 3: This embodiment differs from Embodiment 1 in that the composition of the complete medium for mesenchymal stem cells is: DMEM / F12 basal medium + 10% FBS + 1% glutamine. Others are the same as in the first embodiment.

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Abstract

The invention provides a method for preparing dental pulp mesenchymal stem cells and establishing a dental pulp mesenchymal stem cell bank and a cell resuscitation method, relating to a method for preparing a stem cell bank and a cell resuscitation method. The method comprises the following steps: opening the dental pulp cavity, carrying out primary culture on the dental pulp tissue by a pancreatin substitute half-digestion process, carrying out subculture to obtain dental pulp mesenchymal stem cells, carrying out biological nature detection on the prepared dental pulp mesenchymal stem cells, carrying out amplification culture, cryopreserving, and establishing the dental pulp stem cell bank. The pancreatin substitute half-digestion process for digesting the dental pulp tissue is different from the traditional digestion process; the pancreatin substitute is low in price and easy to purchase; and the half-digestion process effectively prevents the cells from loss and enhances the cryopreservation quantity of the cells. By using teeth as the tissue material source for preparing the mesenchymal stem cells, the invention provides opportunities for preserving dental mesenchymal stem cells.

Description

technical field [0001] The invention relates to a method for preparing a stem cell bank and a cell recovery method. Background technique [0002] Stem cells are a type of cells with self-replication and differentiation potential, which can be differentiated into various functional cells under certain conditions. According to the developmental stage of stem cells, they are divided into embryonic stem cells and adult stem cells. Embryonic stem cells are highly undifferentiated cells that are totipotent in development and can differentiate into all tissues and organs of adult animals, including germ cells. Adult stem cells are undifferentiated cells present in an already differentiated tissue that are capable of self-renewal and specialization to form cells of that type of tissue. Mesenchymal stem cells are adult stem cells with self-replication ability and multilineage differentiation potential. Under specific induction conditions, this kind of stem cells can differentiate ...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A01N1/02
Inventor 芦慧颖张怡王春梅刘艳青
Owner 天晴干细胞股份有限公司
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