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Cartilage stem cell preparation, and preparation method and application thereof

A kind of stem cell preparation and stem cell technology, which can be applied to bone diseases, medical formulas, medical preparations containing active ingredients, etc., can solve the problems of difficult cell proliferation and low cell survival rate, achieve small damage, repair articular cartilage damage, prevent churn effect

Inactive Publication Date: 2017-01-11
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the direct treatment of articular cartilage injury by stem cell injection has the disadvantages of low cell survival rate and difficulty in cell proliferation.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1 Isolation and cultivation of cartilage stem cells

[0040] ①Separate the articular cartilage at the non-stressed part under sterile conditions, peel off the fascia and perichondrium that wrap the cartilage tissue, and put it into a petri dish filled with PBS buffer.

[0041] ② Cut the isolated cartilage tissue into 0.3-0.5mm tissue pieces, transfer them into T25 culture flasks, wash the cartilage with PBS buffer containing double antibody for 3 times.

[0042] ③ Add 0.25% trypsin that is 19-15 times the volume of cartilage, and digest at 37°C for 1-2 hours to stop the digestion.

[0043] ④ Add 0.02% type II collagenase and digest at 37°C for 17-20 hours.

[0044] ⑤ Observe under an inverted microscope, when a single cell is dissociated, add DMEM / F12 complete medium to dilute type II collagenase to stop digestion.

[0045] ⑥Pipe the cell mass into a single cell, filter it with a 200-mesh filter, collect the filtrate, and centrifuge at 1500rpm / min for 5min....

Embodiment 2

[0049] The preparation of embodiment 2 PRP

[0050] ① Sterile collection to obtain 10ml of autologous peripheral whole blood;

[0051] ② Transfer to a 15ml centrifuge tube and centrifuge at 400g for 10 minutes. At this time, it is divided into 3 layers, the upper layer is platelet-poor plasma, the middle layer is platelet-rich, and the lower layer is blood cells;

[0052]③Transfer the upper layer and all the middle layer to a new 15ml centrifuge tube, and centrifuge at 400g for 5 minutes. At this time, it is divided into 3 layers, the upper layer is platelet-poor plasma, the middle layer is rich in platelets, and the lower layer has a small amount of blood cells;

[0053] ④Transfer the upper layer and the middle layer into a new 15ml centrifuge tube, try not to absorb blood cells, centrifuge at 1200g for 5min, collect the required volume of plasma, and resuspend the platelets at the bottom to obtain PRP.

Embodiment 3

[0054] The preparation of embodiment 3 injection preparations

[0055] Cartilage stem cells obtained in Example 1 were resuspended with PRP obtained in Example 2 to 1×10 7 cells / ml for injection into the affected area. The pH of the cell preparation is 7.2; the osmotic pressure range is between 303 mOsm / L.

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Abstract

The invention relates to the technical field of a cell preparation and especially relates to a cartilage stem cell preparation, and a preparation method and application thereof. The cartilage stem cell preparation comprises cartilage stem cells and platelet-rich plasma. The cartilage stem cell preparation provided by the invention can effectively repair joint cartilage injury and has an effect better than the therapeutic effect of sodium hyaluronate combined with mesenchymal stem cells.

Description

technical field [0001] The invention relates to the technical field of cell preparations, in particular to a cartilage stem cell preparation and its preparation method and application. Background technique [0002] Articular cartilage is a viscoelastic substance, and different active parts exhibit different load-bearing characteristics. This important characteristic of articular cartilage and its function of reducing joint surface friction is the basic function of cartilage ultrastructural composition and chemical composition. Articular cartilage is composed of sparse cells without blood vessels, lymph and nerve supply. Articular cartilage plays an important role in joint movement, and its structure is very fine and scientific to meet different functional needs. Trauma or sports injury often leads to acute injury of articular cartilage, causing joint pain, swelling and dysfunction, seriously affecting the quality of life of patients. [0003] At present, the treatment meth...

Claims

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Application Information

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IPC IPC(8): A61K35/32A61P19/02A61K35/16A61K35/19
CPCA61K9/0019A61K35/16A61K35/19A61K35/32A61K2300/00
Inventor 陈海佳葛啸虎王一飞刘帅王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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