Skin ulcer healing combination and preparation method thereof
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
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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...
Embodiment 2
[0045] Example 2 Preparation of umbilical cord mesenchymal stem cell supernatant freeze-dried powder
[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.
Embodiment 3
[0049] Example 3 Preparation of platelet-rich plasma
[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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