Method for preparing stem cell active factor
An active factor and stem cell technology, applied in the field of stem cells, can solve problems such as hindering popularization and application, secondary pollution of stem cells, dry skin, etc.
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Embodiment 1
[0087] Example 1: Preparation of Stem Cell Active Factor Concentrate
[0088] (1) Acquisition of umbilical cord-derived mesenchymal stem cells:
[0089]The umbilical cord of a healthy full-term cesarean section was taken, and prepared by separating Wharton's Jelly (Wharton's Jelly) and using the Wharton's Jelly suspension method; put Wharton's Jelly in a T75 culture bottle, and add an appropriate amount of mesenchymal stem cells (MSC ) Serum-free medium, cultured in a 37°C, 5% CO2 incubator, cell colonies appear after 7 days of culture; cells reach saturation in about 12 days, and can be subcultured; the cells are long spindle-shaped, which is a typical fibroblast Cell-like morphology (see for example figure 1 );
[0090] (2) Cell expansion:
[0091] When the cell confluency reaches 70-80%, gently wash the cells in the culture flask with physiological saline 1-2 times, add 4ml of digestion solution (0.25% trypsin), and leave it at room temperature for 1-2 minutes. Observe...
Embodiment 2
[0100] Embodiment 2: Preparation of stem cell active factor concentrate
[0101] (1) Acquisition of umbilical cord-derived mesenchymal stem cells:
[0102] The umbilical cord of a healthy full-term cesarean section was taken, and prepared by separating Wharton's Jelly (Wharton's Jelly) and using the Wharton's Jelly suspension method; put Wharton's Jelly in a T75 culture bottle, and add an appropriate amount of mesenchymal stem cells (MSC ) Serum-free medium, cultured in 37°C, 5% CO2 incubator, cell colonies appear after 7 days of culture; cells reach saturation in about 10 days, and can be subcultured; cells are long spindle-shaped, which is a typical fibroblast cell-like morphology;
[0103] (2) Cell expansion:
[0104] When the cell confluency reaches 70-80%, gently wash the cells in the culture flask with physiological saline for 1-2 times, add 5ml of digestive solution (0.25% trypsin), and leave it at room temperature for 1-2 minutes. Observe under the microscope that ...
Embodiment 3
[0112] Embodiment 3: Preparation of stem cell active factor concentrate
[0113] (1) Acquisition of umbilical cord-derived mesenchymal stem cells:
[0114] The umbilical cord of a healthy full-term cesarean section was taken, and prepared by separating Wharton's Jelly (Wharton's Jelly) and using the Wharton's Jelly suspension method; put Wharton's Jelly in a T75 culture bottle, and add an appropriate amount of mesenchymal stem cells (MSC ) serum-free medium, cultured in a 37°C, 5% CO2 incubator, and cell colonies appeared after 7 days of culture; the cells reached saturation in about 13 days and could be subcultured; the cells were long spindle-shaped, which is a typical fibroblast cell-like morphology;
[0115] (2) Cell expansion:
[0116] When the cell confluence reaches 70-80%, gently wash the cells in the culture flask with physiological saline for 1-2 times, add 3ml of digestion solution (0.25% trypsin), and leave it at room temperature for 1-2 minutes. Observe under ...
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