Mesenchymal stem cell preparation for repairing articular cartilage injury or coloboma, preparation method and application thereof
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A technology for stem cell and cartilage damage, applied in the direction of bone/connective tissue cells, animal cells, vertebrate cells, etc., can solve the problems of weakened cell differentiation function state, cell concentration and quality affecting repair effect, etc., to achieve the effect of promoting repair
Active Publication Date: 2018-11-23
马琳 +1
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Among them, injecting directly through the joint cavity is a simple and easy operation method, but the directly injected MSCs may cause the number of transplanted cells to decrease continuously and the state of cell differentiation function to weaken
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Embodiment 1
[0030] Preliminary Example 1 Isolation and Culture of Synoviocytes Used in Co-Culture
[0031] 1) Screen patients without immune diseases such as rheumatism, rheumatoid and osteoarthritis, and collect healthy synovial tissue from the knee joint of patients with meniscus or cruciate ligament injury or synovial donors through arthroscopic surgery, and transfer the tissue to Stored in PBS buffer containing 1% double antibody.
[0032] 2) Under aseptic conditions, the synovium tissue was placed in a petri dish and shredded, washed 3 times with PBS buffer solution, and collected into a centrifuge tube.
[0033] 3) Add 4% type I collagenase 5 times the volume of the tissue, place it in a constant temperature shaker, and digest at 150 rpm at 37°C for 2-3 hours.
[0034] 4) After the tissue block was completely digested, the cell suspension was filtered through a 70 μm cell strainer, and the collected filtrate was centrifuged at 1500 rpm for 5 min.
[0035] 5) Discard the supernatan...
Embodiment 2
[0057] The preparation of embodiment 2 preparations
[0058] The 11 groups of co-cultured umbilical cord mesenchymal stem cells cryopreserved in Example 1 were treated as follows: resuscitated in a 37°C water bath, washed once with normal saline, centrifuged at 2000rpm for 5min, and the supernatant was discarded.
[0059] 1) Resuspend the cells with normal saline as an adjuvant, and adjust the cell concentration to 1×10 6 / mL, 1mL each, inhaled into a syringe for use.
[0060] 2) Resuspend the cells with physiological saline as an adjuvant, and adjust the cell concentration to 10×10 6 / mL, 1mL each, inhaled into a syringe for use.
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Abstract
The invention discloses a mesenchymal stem cell preparation for repairing articular cartilage injury or coloboma, a preparation method and application thereof. According to the preparation method, mesenchymal stem cells and synovial cells are indirectly cocultured, so as to obtain the mesenchymal stem cells after coculture, and then the mesenchymal stem cells are cryopreserved, umbilical cord mesenchymal stem cells, which are acquired after the synovial cells and the umbilical cord mesenchymal stem cells are inoculated at density ratio of 4:1 and cocultured, are easier to differentiate towardcartilage direction under a same induction condition. The preparation method utilizes a constructed animal knee joint cartilage injury or coloboma model to prove that the umbilical cord mesenchymal stem cells cocultured with the synovial cells have an obvious effect of facilitating the repairing of articular cartilage injury or coloboma. The invention further provides a preparation which is prepared from the cocultured mesenchymal stem cells. The preparation can be applied to clinic repairing of articular cartilage injury or coloboma; the preparation can effectively promote the repairing of articular cartilage injury or coloboma through anticular injection; the therapeutic effect of the preparation is obviously better than that of a pure mesenchymal stem cell preparation.
Description
technical field [0001] The present invention relates to a preparation for repairing articular cartilage damage / defect, in particular to a mesenchymal stem cell preparation for repairing articular cartilage damage / defect and a preparation method thereof, and the present invention further relates to the medicinal use of the mesenchymal stem cell preparation, namely repairing articular cartilage The damage / defect belongs to the field of mesenchymal stem cell preparation and its preparation and application. Background technique [0002] Articular cartilage is a layer of hyaline cartilage covering the joint surface, composed of chondrocytes and matrix. The surface of articular cartilage is smooth, which can reduce friction, protect joints from wear and tear while bearing sports mechanical loads, and play an important role in joint activities. The main causes of articular cartilage damage / defect include long-term heavy and high-load exercise and middle-aged and elderly osteoarthr...
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