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Skin ulcer healing combination and preparation method thereof
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A composition and umbilical cord technology, applied in the field of medicine, can solve the problems of blood dilution, low stem cell homing rate, and inability to play a role
Inactive Publication Date: 2016-11-09
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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However, the lysed components of PRP will stay in various parts of the body after intravenous reinfusion, and only some will return to the injured site, and will be diluted by the blood and cannot play a role
Moreover, relying solely on the intravenous infusion of stem cells, the homing rate of stem cells is low, the effect is low, and the repairing effect on the ulcer site is low.
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Embodiment 1
[0038] In order to further understand the present invention, the present invention will be described in detail below in conjunction with specific examples. Example 1 Separation and culture of umbilical cord mesenchymal stem cells:
[0039] 1. Take one umbilical cord of a healthy pregnant woman and wash it with PBS+200U / ml double antibody solution (penicillin and streptomycin) for 3 times;
[0040] 2. Cut the umbilical cord into 2cm long pieces, strip off the arterial and venous tubes and adventitia, and cut the middle Huarenheit into pieces of 1-2mm 2 Small pieces, evenly attached to a 10cm Petri dish, 37°C 5% CO 2 Put it upside down in the incubator for 1-2 hours;
[0041] 3. Add 5ml of complete medium (X-VIVO15 serum-free medium + 10ng / ml EGF), change the medium in half after 2-3 days, and culture for 6-8 days. After the cells climb out of the tissue piece, discard the tissue piece Continue to cultivate.
[0042]4. After growing to 80% confluence, discard the culture med...
[0046] 1. Collect the supernatants of P2 and P3 generation umbilical cord mesenchymal stem cells, and filter through a 0.45 μm filter membrane to remove large particles;
[0047] 2. The filtrate is then filtered through a 0.22 μm filter membrane to remove small particles below 3KDa, and the two filter membranes are circulated for 3 hours, concentrated using a Slice200 tangential flow device, and the concentrated solution is collected;
[0048] 3. The concentrated solution is made into a freeze-dried powder by a freeze dryer for later use.
[0050] 1. Take 20ml of peripheral blood, transfer to a 50ml centrifuge tube, centrifuge at 200-400g for 5-10min, and take 8-10ml of upper plasma;
[0051] 2. Collect plasma, centrifuge at 200g-400g for 5-10min, and collect the upper plasma;
[0052] 3. Centrifuge again at 1200g-2000g for 5-10min, absorb the upper layer of plasma and discard it, keep the bottom sediment and 1ml of plasma, and resuspend to obtain PRP (platelet-rich plasma).
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Abstract
The invention belongs to the field of medicines, and particularly relates to a skinulcer healing combination and a preparation method thereof. The combination comprises an intravenous drip component and an outward spray component, wherein the intravenous drip component is normal saline containing umbilical cord mesenchymal stem cells, and the outward spray component comprises umbilical cordmesenchymal stem cell supernate freeze-dried powder, platelet rich plasma, a stromal cell-derived factor-1 and normal saline. The umbilical cord mesenchymal stem cells are intravenously re-infused, various factors in umbilical cord mesenchymal stem cell culture solution are mixed with the platelet rich plasma and the stromal cell-derived factor-1 and locally sprayed outwards, and rapider and better skinulcer healing effects are achieved. Experiments indicate that the skinulcer healing effect of the combination is obviously superior to that of the prior art.
Description
technical field [0001] The invention belongs to the field of medicine, and in particular relates to a composition for repairing skin ulcers and a preparation method thereof. Background technique [0002] Skin ulcer is a common and frequently-occurring disease in traditional Chinese medicinesurgery. It is characterized by skin ulcer as the main clinical manifestation and long-term non-healing. Skintissue defect is liquefied, infected and necrotic. Localized skin tissue defects caused by neurological dysfunction, abnormal immune function or congenital skin defects. Because the sore surface is difficult to heal and consumes a lot, it causes great psychological pressure and economic loss to the patient, and seriously affects the patient's health and quality of life. [0003] Timely and effective wound repair is the core of treating ulcer disease. In recent years, with the in-depth research on stem cells, the role of stem cells in the repair of skin ulcer wounds has also been...
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