Cell preparation for treating osteoarthritis and preparation method thereof
A technology of cell preparations and osteoarthritis, which is applied in the field of cell preparations for the treatment of osteoarthritis and its preparation, can solve the problems of inability to give full play to the biological properties of cells, unfavorable recovery from arthritis, and single cell types, so as to achieve relief and Injury repair, improvement of joint cavity microenvironment, clear and obvious effect of tissue structure
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[0048] The preparation of the above-mentioned cell preparation for the treatment of osteoarthritis includes: isolation and culture of mesenchymal stem cells; induction of differentiation of mesenchymal stem cells; preparation of platelet-rich plasma; preparation of cell preparations. The preparation of preparation, concrete preparation process is:
[0049] Step 1: Isolation and culture of mesenchymal stem cells:
[0050] The selected seed cells include mesenchymal stem cells from various sources, the mesenchymal stem cells can be bone marrow mesenchymal stem cells, fat mesenchymal stem cells, peripheral blood mesenchymal stem cells, synovial mesenchymal stem cells, umbilical cord mesenchymal stem cells For any one of stem cells and umbilical cord blood mesenchymal stem cells, the culture method of seed cells and the cell culture medium used can refer to relevant literature (Hu Bin, Liu Xiaohua, Zhao Lin. Isolation, Culture and Identification of Bone Marrow Mesenchymal Stem Cel...
Embodiment 1
[0060] Example 1: Adipose-derived mesenchymal stem cells as seed cells
[0061] Step 1. Isolation and culture of adipose-derived mesenchymal stem cells
[0062] New Zealand rabbits were routinely anesthetized, adipose tissue was obtained subcutaneously from the abdomen, washed three times with PBS solution, blood vessels and connective tissue were removed, cut into pieces, added 3 times the volume of type I collagenase, digested at 37°C for 2 h, and filtered through a 200-mesh filter Filter out the undigested large adipose tissue, centrifuge to discard the supernatant, resuspend the cells with PBS, wash 3 times, then add 10% FBS-containing low-sugar DMEM to resuspend the pelleted cells, inoculate on a plate, and store at 37°C with volume fraction 0.05% CO 2 Cultivate in an incubator, change the medium every other day, and digest and passage with 0.25% trypsin when the cells cover 80% of the bottom of the bottle.
[0063] Step 2. Induction and differentiation of adipose-deriv...
Embodiment 2
[0071] Example 2: Human umbilical cord mesenchymal stem cells are used as seed cells, and the umbilical cord is derived from donor donation.
[0072] Step 1. Isolation and culture of umbilical cord mesenchymal stem cells:
[0073] Take the umbilical cord tissue out of the collection bottle, wash it twice in alcohol and transfer it to sterile PBS to remove connective tissue, congestion, etc., and then wash it thoroughly to remove blood stains. Transfer the umbilical cord into a large Petri dish, cut it into 3-4cm lengths and then separate it longitudinally, so that the umbilical cord tissue changes from tubular to flake. The umbilical cord tissue is further separated and cut into 2-3mm 3 Tissue pieces of different sizes were planted one by one in a petri dish pre-coated with D / F12 medium containing 20% fetal bovine serum, and placed at 37°C with a volume fraction of 5% CO 2 In the saturated humidity incubator, add 0.5ml of medium every 24 hours, change the medium after 72 h...
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