Umbilical cord-derived mesenchymal stem cells as well as preparation method and application thereof
A mesenchymal stem cell source technology, applied in the field of stem cells, can solve the problem of mesenchymal stem cells being scarce
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[0061] The preparation method of the above-mentioned umbilical cord-derived mesenchymal stem cells uses umbilical cord tissues other than venous blood vessels and arterial blood vessels as raw materials, adopts the tissue block adhesion method to prepare mesenchymal stem cells, simplifies the separation steps, and is convenient for existing manual operations to be transformed into Automated equipment, which greatly improves the utilization rate of the umbilical cord, makes efficient use of limited samples, and greatly increases the yield of mesenchymal stem cells. Importantly, this study unexpectedly found that the cell mass obtained from all umbilical cord tissue after removal of venous and arterial vessels was higher than that obtained from only Wharton's jelly or only amniotic membrane when the size of the tissue block was the same of cells. Moreover, it has been verified that the mesenchymal stem cells prepared according to the above method for preparing the umbilical cord...
Embodiment 1
[0069] 1. Umbilical cord collection: collect full-term healthy cesarean section fetal umbilical cords, immerse in PBS containing 1% (m / v) penicillin and 1% (m / v) streptomycin, and place on ice.
[0070] 2. Tissue separation: cut the umbilical cord into small sections about 3 cm long in the ultra-clean bench, cut them longitudinally, rinse them repeatedly with sterile PBS until the liquid is free of blood, and remove one venous vessel and two arterial vessels with a hemostat and an ophthalmologist (three in total), to get the umbilical cord without arterial blood vessel and venous blood vessel, to remove the umbilical cord of arterial blood vessel and venous blood vessel such as figure 1 shown.
[0071] 3. Preparation of adherent tissue block: cut the umbilical cord obtained in step 2 with arterial and venous vessels removed to less than 2mm 3 Small pieces, rinsed with PBS and drained, and placed in the incubator for 30 minutes, then added DMEM / F12 medium containing 15% (v / v) ...
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