A stem cell extract that eliminates stretch marks on the skin
A stem cell and extract technology, applied in the field of stem cells, can solve problems such as insufficient effect
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Embodiment 1
[0027] The isolation and expansion culture of placental stem cells are as follows:
[0028] (1) Choose normal human placenta, cut the placenta tissue into small pieces, fully wash away the free cells with cold phosphate buffered saline (PBS), then use 0.25% trypsin / HBSS to vibrate and digest twice in a water bath at 37°C, each time For 10 minutes each time, the digested solution was collected twice and neutralized with an equal amount of DMEM containing 10% human serum. After centrifugation, use 10% human serum DMEM to suspend the cells and plant them on the cell culture dish. After 24 hours, change the medium to remove the suspended cells, and then change the medium every 3 days. Stem cell colonies can be seen in 7-10 days, and continue to culture until the cells in the culture dish Cells were harvested by trypsinization at 70% saturation.
[0029] (2) Sorting CD140a positive cells by immunomagnetic bead method, the basic steps are as follows (for details, please refer to th...
Embodiment 2
[0035] The isolation, expansion and culture of umbilical cord stem cells are as follows:
[0036] (1) Separation of umbilical cord cells by adherent method: wash the normal human umbilical cord repeatedly with PBS, peel off the blood vessels and adventitia, collect Wharton's jelly, cut into small pieces of about 1 mm, and suspend to contain 10% human serum Inoculate cell culture dishes in DMEM in an amount that does not suspend tissue pieces at 37 °C and 5% CO 2 Cultivate in an incubator, carefully change the medium after 3-5 days, and cells grow from the periphery of the tissue after 5-7 days, digest with trypsin and collect the cells.
[0037] (2) Sorting CD140a positive cells by immunomagnetic bead method, the steps are the same as the step (2) of the isolation and expansion culture of placental stem cells in Example 1.
[0038] (3) Cultivate umbilical cord stem cells with serum-free medium, the steps are the same as the step (3) of the isolation and expansion culture of p...
Embodiment 3
[0040] Preparation and Application of Stem Cell Extract
[0041] (1) Preparation of keratinocyte conditioned medium: Human foreskin-derived keratinocytes were cultured in full K-SFM medium (Life Technologies) to 70% saturation, and replaced with additive-free K-SFM medium for 24 hours. hours, collect the culture medium, centrifuge at 4°C and 2095G for 30 minutes, discard the bottom sediment to collect the supernatant, and store it at -80°C for future use.
[0042] (2) Placental stem cells were incubated with 10% keratinocyte conditioned medium and 90% MesenGro serum-free medium at 37°C and 5% CO 2 Cultured in the incubator for 3 days to 80-90% saturation, trypsinized and passaged.
[0043] (3) The above-passaged cells were kept in 10% keratinocyte conditioned medium and 90% MesenGro serum-free medium at 37°C and 5% CO 2 Cultivate in the incubator for 2 days to 70% saturation, remove the original medium, add PBS or DMEM equivalent to half the amount of the original medium, co...
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