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Method for inducing dental pulp mesenchymal stem cells to be differentiated into nerve cells

A technology for stem cells and nerve cells, which is applied in the field of inducing the differentiation of dental pulp mesenchymal stem cells into nerve cells, which can solve the difficulty of purifying the final product of cell culture, the contamination of exogenous viruses and pathogenic factors, and the allergic reaction of xenogeneic serum, etc. problems, to facilitate the timing of treatment, improve biosafety, and increase the rate of differentiation

Active Publication Date: 2015-06-24
中国医科大学 +1
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Problems solved by technology

However, due to the complex composition of fetal bovine serum, the use of serum-containing medium to culture cells has potential cytotoxicity, foreign virus and pathogenic factor contamination, which makes the standardization of cell culture and the purification of final products more difficult, and the residue of heterogeneous serum clinically allergic reactions

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  • Method for inducing dental pulp mesenchymal stem cells to be differentiated into nerve cells
  • Method for inducing dental pulp mesenchymal stem cells to be differentiated into nerve cells
  • Method for inducing dental pulp mesenchymal stem cells to be differentiated into nerve cells

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Embodiment Construction

[0042] The present invention will be further described in detail below in conjunction with specific examples.

[0043] The invention provides a method for inducing the differentiation of dental pulp mesenchymal stem cells into nerve cells, comprising the following steps.

[0044] Step 1, extraction of amniotic membrane stem cells and collection of amniotic membrane stem cell culture supernatant.

[0045] (1) Placenta collection: With the informed consent of the puerpera, the placenta was collected from healthy fetuses delivered by cesarean section at term, stored at 4°C, and transported to the laboratory within 24 hours. Maternal blood should be collected within 7 days before delivery for AIDS testing, hepatitis B testing, hepatitis C testing, cytomegalovirus testing, syphilis testing, alanine aminotransferase testing, and mycoplasma testing.

[0046] (2) Primary extraction of amniotic membrane stem cells: Bluntly separate the amniotic membrane from the umbilical cord surface...

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Abstract

The invention provides a method for inducing dental pulp mesenchymal stem cells to be differentiated into nerve cells. The method comprises the following steps: extracting the dental pulp mesenchymal stem cells by virtue of an enzyme digestion method, inoculating the dental pulp mesenchymal stem cells to a culture dish coated with polylysine and laminin, adding an optimized culture medium, and performing culture amplification under a low-oxygen condition; then performing double-layer coating on the collected dental pulp mesenchymal stem cells by using an acellular amniotic membrane substrate, and performing induced differentiation by using a nerve cell induction solution; and finally, maintaining long-term culture of the nerve cells by using a nerve cell maintenance solution. According to the method provided by the invention, the optimized culture medium and the low-oxygen condition are used for culture, so that the number of the dental pulp mesenchymal stem cells is increased; the double-layer acellular amniotic membrane substrate is used for simulating a neurolemma structure to achieve an effect of guiding the growth of the nerve cells, and meanwhile, the nerve cell induction solution is used for inducing the dental pulp mesenchymal stem cells to be differentiated towards nerves, so that the differentiation rate is improved; and the nerve cell maintenance solution is used for maintaining the activity of the nerve cells for a long term, so that a clinician can flexibly control the treatment time conveniently.

Description

technical field [0001] The invention relates to the field of cell culture and induced differentiation of stem cells, in particular to a method for inducing the differentiation of dental pulp mesenchymal stem cells into nerve cells. Background technique [0002] Stem cells are primitive cells with certain self-renewal ability and multi-directional differentiation potential in a suitable microenvironment, which can differentiate into various tissue cells such as fat, cartilage, bone, muscle, tendon, nerve, liver, and cardiac muscle. Mesenchymal stem cells (mesenchymal stem cells, MSCs) are non-terminal differentiated cells derived from mesoderm, which have the characteristics of mesenchymal cells, endothelial cells and epithelial cells, and are an important source of seed cells for cell therapy and tissue engineering . In addition to bone marrow, peripheral blood, fat, and umbilical cord, human dental pulp is also an important source of mesenchymal stem cells. [0003] Denta...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/079
Inventor 于艳秋宋冰吕亚楠荣耀星邓雯雯时兆田张停停李秀文
Owner 中国医科大学
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