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Use of mesenchymal stem cells of dental pulp in obtaining of artificial spinal cord and preparation method for artificial spinal cord

A technique for stem cells and dental pulp, which is applied to the use of dental pulp mesenchymal stem cells to obtain artificial spinal cords and the field of preparation thereof, which can solve the problems of time-consuming and labor-intensive extraction of exosomes.

Pending Publication Date: 2021-05-18
SECOND AFFILIATED HOSPITAL OF COLLEGE OF MEDICINEOF XIAN JIAOTONG UNIV
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Problems solved by technology

[0005] The purpose of the present invention is to provide the use of dental pulp mesenchymal stem cells to obtain artificial spinal cord and its preparation method to solve the problem of time-consuming and labor-consuming exosome extraction process for the treatment of spinal cord injury

Method used

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  • Use of mesenchymal stem cells of dental pulp in obtaining of artificial spinal cord and preparation method for artificial spinal cord
  • Use of mesenchymal stem cells of dental pulp in obtaining of artificial spinal cord and preparation method for artificial spinal cord
  • Use of mesenchymal stem cells of dental pulp in obtaining of artificial spinal cord and preparation method for artificial spinal cord

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Embodiment 1

[0049] 1. Cell Culture

[0050] Human dental pulp stem cells (DPSCs) were isolated and cultured. Healthy deciduous teeth extracted for orthodontic reasons (a stomatological hospital) were collected as tooth samples. Digestion was performed with type I collagenase (3 mg / mL; Sigma, USA) at 37°C for 1 hour. The single cell suspension was 1×10 5 cells / cm 2 The density is inoculated in a 75cm 2 in the culture flask. After 24 hours, wash and discard unattached cells. Adherent cells were resuspended and cultured in a medium containing 10% fetal bovine serum (Hyclone, Logan, Utah, USA), 2mmol / L glutamine (Invitrogen, Carlsbad, CA, USA), 100U / mL penicillin (Invitrogen, Carlsbad, CA). , USA) and 100 mg / mL streptomycin (Invitrogen, Carlsbad, CA, USA) in α-MEM (Gibco, USA) medium for 2 weeks.

[0051] 2. Cell Identification

[0052] In order to identify cell surface markers of dental pulp mesenchymal stem cells, 3×10 5 Individual cells were incubated with PE or FITC-conjugated mo...

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Abstract

The invention discloses use of mesenchymal stem cells of dental pulp in obtaining of an artificial spinal cord and a preparation method for the artificial spinal cord. The preparation method consists of the following steps: (1) acquiring healthy deciduous teeth removed due to deformity correcting, digesting the healthy deciduous teeth with type 1 collagenase, inoculating a culture flask with the digested substance, carrying out washing to discard nonadherent cells, re-suspending adherent cells, then, carrying out culture, so as to obtain the mesenchymal stem cells of the dental pulp; (2) culturing the mesenchymal stem cells of the dental pulp with an alpha-MEM culture medium, so as to propagate the mesenchymal stem cells of the dental pulp; and (3) selecting third-generation dental-pulp mesenchymal stem cells, culturing the dental-pulp mesenchymal stem cells with an alpha-MEM culture medium containing vitamin C when the cell fusion rate is 90%so as to obtain a white membrane-shaped structure, and carrying out in-vitro molding, thereby finally obtaining the artificial spinal cord with a three-dimensional structure. The artificial spinal cord obtained by the preparation method can be used for inhibiting cicatrization and inflammation and promoting axon regeneration; and among rats suffering from complete spinal injury, Spinor therapy causes remarkable movement improvement, perception recovery and faster urination reflection recovery.

Description

technical field [0001] The invention belongs to the field of preparation methods of artificial spinal cord, and in particular relates to the use of dental pulp mesenchymal stem cells to obtain artificial spinal cord and its preparation method. Background technique [0002] Spinal cord injury (SCI) is a complex pathophysiological cascade reaction, especially involving neurological deficits caused by spinal cord injury, with an annual incidence of more than one in a thousand worldwide. Worldwide, more than 700,000 new cases of SCI are diagnosed each year due to mechanical injuries of the spine. In the past decade, the development and popularization of surgical technology has caused the number of SCI deaths to drop by more than 35%. However, surgery cannot reverse this permanent disability due to the poor innate ability to regenerate neurons in the adult CNS. Even if alive, SCI survivors, especially those with severe or even complete SCI, suffer extreme physical suffering and...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A61K35/28A61P25/28
CPCC12N5/0664A61K35/28A61P25/28C12N2513/00C12N2509/00
Inventor 刘文佳何旺骁闫瑾
Owner SECOND AFFILIATED HOSPITAL OF COLLEGE OF MEDICINEOF XIAN JIAOTONG UNIV
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