Hydro lifting instrument injection umbilical cord mesenchymal stem cell preparation and application thereof
A stem cell and stem cell technology, applied in animal cells, vertebrate cells, cell dissociation methods, etc., can solve problems accompanied by pain and bruises, sequelae, and treat symptoms rather than the root cause, so as to improve pathological conditions and increase blood content. The amount of water, the effect is remarkable and long-lasting effect
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Embodiment 1
[0041] This example provides a method for establishing a safe, effective, and sufficient number of cell banks. The cells in the cell bank are certified by a professional third party, and the amount of cells can meet the continuous supply of clinical treatment.
[0042] The umbilical cord tissue is obtained after the newborn is born, and the donor must undergo a strict physical examination, including various virus tests and infectious disease tests, and sign an informed consent form and a donation agreement. In the sterile operating room, the umbilical cord freshly separated from the human body is quickly transferred to 4°C sterile sodium chloride injection, sealed, and transported to a professional cell preparation center through a cold chain within 4 hours.
[0043] In this embodiment, the specific steps for establishing the cell bank are as follows:
[0044] (1) Transfer the umbilical cord tissue to a pharmaceutical-grade GMP workshop, undergo strict treatment, sterilize wit...
Embodiment 2
[0054] This example provides a method for preparing clinical-grade umbilical cord mesenchymal stem cells using a cell factory, and the specific steps are as follows:
[0055] (1) Take 1 tube from the stem cell seed bank and pack 5-10×10 6 Cells / ml / tube of P2 generation umbilical cord mesenchymal stem cells, after recovery, press 5~10×10 3 / cm 2 The density is inoculated into a 2-layer cell factory (NEST, Cat No.771101) for stem cell expansion culture, so that the stem cells can be expanded to a certain number (P3 generation), wherein: the source of stem cells inoculated in each 2-layer cell factory For the cells in the same cell cryopreservation tube, the total number of cells inoculated in each 2-layer cell factory is 6.395×10 6 The volume of cell suspension added to each layer is 150-200ml, and the culture conditions are: at 37°C, 5% CO 2 Adhesive culture in an incubator with saturated humidity for 3 to 5 days;
[0056] (2) When the degree of cell confluence reaches abou...
Embodiment 3
[0061] This embodiment provides a method for identifying the umbilical cord mesenchymal stem cells prepared in Example 2, specifically as follows:
[0062] In 2006, the International Society for Cellular Therapy (ISCT) established the identification criteria for mesenchymal stem cells as:
[0063] (1) grow adherently in plastic petri dishes, and proliferate highly in medium containing serum;
[0064] (2) Express cell surface markers CD105, CD73, CD90, but not CD45, CD34, CD14 or CD11b, CD79a or CD19;
[0065] (3) It can differentiate into osteoblasts, adipocytes and chondrocytes in vitro.
[0066] In this example, stem cell phenotype identification is carried out according to this minimum standard.
[0067] After the proliferation and confluence of stem cells reached 90% in the 10-layer cell factory, use the multi-layer cell culture machine observation platform to observe the cell morphology, such as figure 1 As shown, the stem cells grow adherently, spindle-shaped, and are n...
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