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Neuroprotective composition, preparation process thereof and medical uses thereof

A technology of neuroprotection and composition, which is applied in the direction of biochemical equipment and methods, drug combination, nervous system diseases, etc., and can solve problems such as inefficiency

Active Publication Date: 2019-07-05
祐安細胞生醫科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005]Despite the high severity of SAH and its associated complications, current interventions to treat SAH are relatively ineffective

Method used

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  • Neuroprotective composition, preparation process thereof and medical uses thereof
  • Neuroprotective composition, preparation process thereof and medical uses thereof
  • Neuroprotective composition, preparation process thereof and medical uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1: Isolation and culture of dental pulp stem cells (DPSC)

[0053] The upper teeth were harvested intact from 3-week-old male Wistar rats, and then the tissue was extracted using a syringe needle and transferred to a 25 cm 2 flask (Corning, Inc., New York, USA). The pulp was washed twice with phosphate buffered saline and treated with type II collagenase (50-100 U / ml) for 1 hour. The tissue was then centrifuged at 200 rpm for 4 minutes to obtain cell pellets, which were subsequently washed twice with phosphate buffered saline. Supplemented with 10% FBS ( The cells were cultured in Dulbecco's modified Eagle's medium (DMEM) nutrient mixture F12 (DMEM / F12; Life Technologies, Karlsruhe, Germany) in Grand Island, New York, USA) for several days until the cell number reached more than 1 × 10 6 indivual. Cells were trypsinized to detach and washed twice with phosphate-buffered saline, then resuspended in 0.5 ml phosphate-buffered saline. Deplete the cell suspen...

Embodiment 2

[0054] Example 2: Preparation of biologically active media fractions

[0055] Conditioned medium was produced as follows: to 150 cm 2 80% confluent MSCs of 2-5 passages in cell culture flasks (Corning, Inc. New York, USA) were added with 20 mL / bottle of serum-free DMEM / F12 (Life Technologies, Karlsruhe, Germany), and then cultivated for 72 hours. The DMEM / F12 was supplemented with 10 ml of Hank's balanced salt solution (HBSS; Life Technologies, Carlsbad, CA, USA) and 300 μl of penicillin-streptomycin (1%). For the in vitro and in vivo experiments described below, a Tangential Flow Filtration (TFF) system (Millipore Co., St. Charles, MO, USA) equipped with a 5 kDa cut-off membrane (Millipore Co., Billerica, MA, USA) was used Conditioned medium was further concentrated according to the manufacturer's instructions, thereby obtaining medium fractions with nominal molecular weights equal to and less than 5 kDa.

Embodiment 3

[0056] Embodiment 3: rat SAH model

[0057]Male Wistar rats (250 to 300 g) were used in this example. All surgical procedures were performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals, and animal experiments were approved by the local Animal Care and Use Committee. The anesthetic was 2.5% isoflurane with 70% nitrous oxide and 27.5% oxygen. Administration was administered through a tracheostomy tube to ensure deep sedation, which was confirmed by the absence of pain reflexes in the hindlimb and forelimb as well as absence of the corneal reflex. Maintain normal, effortless breathing throughout the procedure. Animals were placed on a heat blanket, temperature was monitored with a rectal probe, and body temperature was maintained at 37°C. Monitor blood pressure and maintain it at 100-120mmHg.

[0058] Animals were randomly divided into three groups, namely: SAH group, in which 0.3 ml of fresh autologous blood was inj...

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Abstract

The invention relates to a neuroprotective composition derived from mesenchymal stem cells, especially a neuroprotective composition derived from the primary culture of dental pulp mesenchymal stem cells. The invention also relates to processes of preparing the neuroprotective composition, and medical uses of the neuroprotective composition in the treatment of neurological diseases associated withneuronal damage, including subarachnoid haemorrhage and Parkinson's disease.

Description

technical field [0001] The present invention relates to a neuroprotective composition derived from mesenchymal stem cells, in particular to a neuroprotective composition derived from the primary culture of dental pulp mesenchymal stem cells, its preparation method, and its application in the treatment of neurons Medical use in injury-related neurological disorders including subarachnoid hemorrhage and Parkinson's disease. Background technique [0002] Subarachnoid hemorrhage (SAH) is the extravasation of blood into the subarachnoid space between the arachnoid and the pia mater surrounding the brain. It occurs in a variety of clinical settings, most commonly head trauma; nontraumatic (or spontaneous) subarachnoid hemorrhage usually occurs in the setting of a ruptured cerebral aneurysm or arteriovenous malformation. Risk factors for spontaneous cases include hypertension, smoking, family history, alcohol and cocaine use. [0003] Aneurysmal SAH carries significant morbidity ...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61P25/00A61P7/04A61P25/16A61P25/28A61P25/14A61P21/00A61P9/10C12N5/0775
CPCA61K35/28A61P25/00A61P7/04A61P25/16A61P25/28A61P25/14A61P21/00A61P9/10C12N5/0664C12N2500/90C12N5/0662
Inventor 陈德福王国川
Owner 祐安細胞生醫科技股份有限公司
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