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Method for preparing, culturing and purifying dental pulp mesenchymal stem cells

A mesenchymal stem cell and cell culture technology, applied in the field of stem cells and regenerative medicine, can solve the problems of inconvenient clinical application of dental pulp mesenchymal stem cells, low cell purity, slow cell migration speed, etc., to facilitate expansion and later clinical application Effect

Inactive Publication Date: 2018-12-07
GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
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Problems solved by technology

[0004] At present, the preparation method of dental pulp mesenchymal stem cells is generally to separate the mesenchymal stem cells by enzymatic digestion or tissue grafting after taking out the dental pulp tissue, but the dental pulp tissue is rich in other tissues such as blood vessels and nerves. The cells obtained by digestion are of low purity and may be mixed with a large number of endothelial cells, fibroblasts and other miscellaneous cells. The transplanted hair cells migrate slowly, and some cells cannot migrate out of the residual tissue block, resulting in cell loss, which brings inconvenience to the clinical application of dental pulp mesenchymal stem cells in the later stage

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  • Method for preparing, culturing and purifying dental pulp mesenchymal stem cells
  • Method for preparing, culturing and purifying dental pulp mesenchymal stem cells
  • Method for preparing, culturing and purifying dental pulp mesenchymal stem cells

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Embodiment Construction

[0024] The technical solutions of the present invention will be described in further detail below in conjunction with specific examples, but the present invention is not limited to the following examples.

[0025] Prepare, cultivate and purify dental pulp mesenchymal stem cells, the steps are as follows:

[0026] 1. The pulp tissue comes from the deciduous teeth that naturally fall out during the process of tooth replacement in children. The stomatological hospital collects samples, and the informed consent is signed before the samples are collected.

[0027] 2. The collected deciduous teeth are placed in sterile tissue protection solution, and transported to the laboratory in a refrigerated environment (4-8°C). The transportation time is controlled within 48 hours. The formula of the tissue protection solution is as follows: H-DMEM (Gibco 12491-015 ), gentamicin sulfate at a final concentration of 25 μg / ml (based on the volume of H-DMEM), and amphotericin B at a final concent...

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Abstract

The invention provides a method for preparing dental pulp mesenchymal stem cells. The method comprises the following steps: adding tissue digestion liquid into dental pulp tissues for digestion; afterthe digestion is finished, adding H-DMEM culture liquid to dilute digestion liquid, filtering tooth fragments, and centrifuging the filtrate to remove supernate, so as to obtain precipitates; cleaning the precipitates with the H-DMEM culture liquid, and carrying out centrifugation again to remove the supernate, so as to obtain the separated dental pulp mesenchymal stem cells. According to the method, the dental pulp mesenchymal stem cells can be effectively prepared, separated and purified.

Description

technical field [0001] The invention belongs to the field of stem cells and regenerative medicine, and in particular relates to a method for preparing, cultivating and purifying dental pulp mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of stem cells with self-replication and multi-directional differentiation capabilities. Under certain conditions, they can differentiate into various functional cells such as bone, cartilage, nerve, and epidermis. They have important applications in the field of regenerative medicine prospect. Mesenchymal stem cells exist in many tissues of the human body, such as common bone marrow, umbilical cord Huatong jelly, umbilical cord blood, placenta and other tissues, but bone marrow-derived mesenchymal stem cells have problems such as difficulty in obtaining materials and causing great harm to the human body; umbilical cord and The mesenchymal stem cells in the placenta and other tissues have the ad...

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0664C12N2500/90C12N2501/119C12N2501/12C12N2501/999C12N2509/00
Inventor 朱建斌江嘉豪何美弟陈炽峰康斯敏陈碧莹李静静蔡祥胜王明珠王进辉
Owner GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
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