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Application of neural stem cells combined with umbilical cord mesenchymal stem cells in spinal cord injury

A technology for neural stem cells and spinal cord injury, which is applied in the direction of nervous system cells, animal cells, and nervous system diseases to achieve the effect of promoting tissue repair, reducing the area, and promoting the repair of nerve damage.

Pending Publication Date: 2021-11-30
HEBEI MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, few studies compared the efficacy of NSC and MSC therapy, but in a subacute SCI model, NSCs and MSCs were transplanted at least 7 days after SCI injury

Method used

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  • Application of neural stem cells combined with umbilical cord mesenchymal stem cells in spinal cord injury
  • Application of neural stem cells combined with umbilical cord mesenchymal stem cells in spinal cord injury
  • Application of neural stem cells combined with umbilical cord mesenchymal stem cells in spinal cord injury

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Experimental program
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Effect test

Embodiment 1

[0033] Ethical statement:

[0034] The collection of all human specimens and the implementation of animal experiments were approved by the Experimental Ethics Committee of Hebei Medical University (guarantee number: 20190505). Experiments were performed in accordance with the Guidelines for the Care and Use of Laboratory Animals.

[0035] Obtain iPSC-derived neural stem cells

[0036] hhiPSCs were generated from dermal fibroblasts using a protocol previously described in a study in our laboratory (cell line: HEBHMUi002-a). Briefly, following the manufacturer's guidelines, use Cytotune TM - iPS 2.0 Sendai Reprogramming Kit (Thermo Fisher Scientific, Waltham, MA, USA), to reprogram fibroblasts at passage 4 into hiPSCs. On day 0 of transduction, 4 reprogramming factors were added to fibroblasts, and the medium was changed after 24 hours to remove virus. On day 7 post-transduction, cells were transferred at the recommended density into 6-well plates coated with Geltrex hESC ce...

Embodiment 2

[0041] Spinal cord injury model establishment

[0042] Eight-week-old BALB / c female immunodeficiency female nude mice (19-22g, n=54) were anesthetized by intraperitoneal injection of 1% sodium pentobarbital (50mg / kg, intraperitoneal injection), Wuhan, Hubei Xinrunde Chemical Co., Ltd. company. China). A laminectomy was performed at the 10th thoracic vertebra to expose the dorsal surface of the spinal dura mater. Spinal cord contusion was induced at T10 level with a Zhongshi impactor (Zhongshi, Beijing, China) weighing 10 g and 5 cm high.

Embodiment 3

[0044] Acquisition of products for the treatment of acute spinal cord injuries in animals;

[0045] The human induced pluripotent stem cell-derived neural stem cells (NSC) combined with human umbilical cord-derived mesenchymal stem cells (MSC) are used in a ratio of 1:1, and the vehicle is PBS;

[0046] The human induced pluripotent stem cells are differentiated into NSCs in vitro through the reprogramming of human peripheral blood mononuclear cells into induced pluripotent stem cells, and the stem cells are differentiated into neural stem cells in vitro under the action of a neural inducer, and placed in a 37°C, 5% CO2 incubator middle;

[0047] In the process of differentiation into neural stem cells, the medium was changed every other day with the neural induction medium preheated at 37°C, co-cultured for seven days, and the cells were digested and passaged on the eighth day to obtain P0 NSCs.

[0048] The human umbilical cord is isolated and cultured as MSC, comprising th...

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Abstract

According to the invention, human induced pluripotent stem cells are differentiated into NSCs in vitro, human umbilical cords are separated and cultured into MSC auxiliary materials, the materials are used according to the proportion of 1: 1, PBS is used as a solvent, an obtained cell biological product is jointly transplanted to the spinal cord of a mouse with acute spinal cord injury (at the thoracic marrow T10 stage) in situ, nerve injury repair is effectively promoted, and the motor function is improved. The product for treating animal acute spinal cord injury can be differentiated into neurons and glial cells in the host body, reduce the glial scar area, reduce fibrosis and reduce the inflammation level, so that the animal motor function, especially limb coordination, is improved, and the product fully plays a role in subsequent treatment of the spinal cord injury and promotion of tissue repair.

Description

technical field [0001] The invention relates to the application of neural stem cells (NSC) derived from human induced pluripotent stem cells combined with mesenchymal stem cells (MSC) derived from human umbilical cord as a product for treating acute spinal cord injury in animals, and belongs to the technical field of combined cell therapy. Background technique [0002] Spinal cord injury (SCI) is a common disease that may lead to destruction and permanent loss of neurological function due to the failure of axon regeneration after injury, thereby interrupting the connection between the brain and the body. Based on available evidence, the incidence of spinal cord injury has increased dramatically in China over the past decade, mainly due to increased spinal trauma from vehicle accidents and falls from heights. The Global Burden of Disease, Injury, and Risk Factors Study reported 0.93 million (95% uncertainty interval [UI] 780,000–1.16 million) new cases of spinal cord injury, ...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61K35/30A61P25/28C12N5/0797C12N5/0775
CPCA61K35/30A61K35/28A61P25/28C12N5/0623C12N5/0668C12N2500/02C12N2509/10A61K2300/00
Inventor 马隽崔慧先孔德胜杜晓峰何晶晶冯宝峰刘博鑫马振环刘鑫郭瑞云张金玉
Owner HEBEI MEDICAL UNIVERSITY
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