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Application of horse fat mesenchymal stem cells in preparing drug for treating horse injury

A technology of mesenchymal stem cells and stem cell preparations, applied in the field of preparation of stem cell preparations, can solve problems such as high recurrence rate, poor prognosis, and long healing period

Pending Publication Date: 2019-05-17
FOSHAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Tendon and ligament injuries are very prevalent in sports horses. Since tendinitis requires a long period of injury recovery, for horse animals, returning to the field often has a poor prognosis. This is also a key factor leading to early retirement of sports horses. Traditional and routine treatment of tendon and ligament injury usually by producing less functional repair tissue, but this healing phase is relatively long and also results in a relatively high recurrence rate

Method used

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  • Application of horse fat mesenchymal stem cells in preparing drug for treating horse injury
  • Application of horse fat mesenchymal stem cells in preparing drug for treating horse injury
  • Application of horse fat mesenchymal stem cells in preparing drug for treating horse injury

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Preparation of horse adipose-derived mesenchymal stem cells

[0036] 1. Isolation and culture of horse adipose-derived mesenchymal stem cells

[0037] Reagents and materials: 0.25% trypsin (0.25g trypsin dissolved in 100ml PBS, purchased from Hyclone Company), 0.1% (g / mL) type I collagenase (Sigma Company), DMEM / F12 basal medium (purchased from Hyclone Company); sterilized tissue shears; 100-mesh cell strainer.

[0038] The "DMEM / F12" basal medium used herein is a basal medium mixed with F12 medium (Ham's F 12nutrient medium) and DMEM medium (Dulbecco's Modified Eagle Medium) at a volume ratio of 1:1.

[0039] The isolation and cultivation steps are as follows:

[0040] (1) Adipose tissue was collected from the neck of the horse. First, the horse was anesthetized, and the neck adipose tissue was aseptically collected. Wash the horse adipose tissue, shake the centrifuge tube to clean the tissue block, and remove impurities such as blood;

[0041] ⑵ Shreddi...

Embodiment 2

[0084] Embodiment 2: Preparation of stem cell preparation

[0085] 1. Components of stem cell preparation: P4 generation adipose-derived mesenchymal stem cells, horse serum albumin, L-glutamine, glucose, red blood cell storage solution.

[0086] 2. Take P4 adipose-derived mesenchymal stem cells from horses in good growth state, and the dose of cell therapy is 5-10×10 6 , on the basis of mesenchymal stem cells every 10 6 Add 0.2ml of stem cell preparation storage solution to each cell, the stem cell storage solution includes: horse serum albumin, L-glutamine, glucose, red blood cell storage solution.

[0087] The proportion of the stem cell preparation storage solution is: 50%wt horse blood albumin, 49%wt red blood cell storage solution, 1%wt L-glutamine, and 1g of glucose is added to each liter of the stem cell preparation storage solution.

Embodiment 3

[0088] Embodiment 3: clinical case treatment

[0089] 1. Injection method

[0090] Tendon injury, with the help of B-ultrasound, one or more injections.

[0091] 2. Amount of Cells to Treat

[0092] 2-10×10 6 Cells (depending on the area of ​​injury), the cells were diluted into 10ml of normal saline when used.

[0093] 3. Course of treatment

[0094] 1-3 injections are a course of treatment, and the intervals are around the first week and the sixth week to observe the recovery after the injection of stem cells, depending on the situation, whether to inject the second or third time, and the cure will take more than 6 months.

[0095] 4. Specific operation

[0096] After shaving, scrub the injured area with iodine and repeat scrubbing with 70% v / v alcohol, inject cells with a 23-gauge needle under the guidance of ultrasound, and resuspend 2-10×10 6 Each cell is slowly injected into the core lesion in a spot or aliquot. Ultrasound guidance is used to ensure that the needl...

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Abstract

The invention provides application of horse fat mesenchymal stem cells in preparing a drug for treating horse injury and also provides a method for isolating and cultivating horse fat mesenchymal stemcells. The application is capable of providing effective stable biologically active horse fat mesenchymal stem cells which can be utilized to effectively treat the horse injury, particularly superficial tendon injury.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a method for separating and culturing horse adipose mesenchymal stem cells and the preparation of stem cell preparations for treating equine tendonitis. Background technique [0002] Overtraining and traumatic tendon and ligament injuries are common in sport horses, and in most cases, tendon and ligament injuries heal (repair) naturally through the formation of scar tissue. However, the natural healing time is long and compared with normal tendon and ligament tissue, the function of scar tissue formed during this repair process is defective, which has an impact on the performance of animal sports and can significantly reduce sports performance. Furthermore, there is a great risk of re-injury. [0003] Mesenchymal stem cells (MSCs) are called pluripotent stem cells. MSCs have a wide range of sources and can be extracted from bone marrow, cartilage, adipose tissue, synovial ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61P21/00A61P19/04C12N5/0775
Inventor 罗惠娜王丙云詹小舒罗冬章
Owner FOSHAN UNIVERSITY
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