Preparation of umbilical cord mesenchymal stem cell excreted factor and application of umbilical cord mesenchymal stem cell excreted factor to hair growth
A technology for stromal stem cells and secreting factors, which is applied in the field of preparation of umbilical cord mesenchymal stem cells secreted factors, can solve problems such as obvious side effects, and achieve the effects of low requirements for instruments and equipment, low cost, and simple preparation methods and steps.
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Embodiment 1
[0057] Embodiment 1. Contains the preparation of multiple polypeptide factor culture medium
[0058] Using StemPro TM Umbilical cord mesenchymal stem cells were cultured in MSC SFM XenoFree medium. When the cells filled the culture flask, they were subcultured to make their confluence 50%-70% within 24 hours. After 24 hours, induction culture was carried out using the induction medium with the specific formulation as follows.
[0059] The specific operation is as follows: remove the subculture medium, wash once with PBS, and add induction medium with different formulations and conditions for induction culture. After culturing for 48 hours, the umbilical cord mesenchymal stem cells can fully secrete polypeptide factors into the medium, and the culture supernatant is collected.
[0060] Four conditions were used for induction culture, and the components of the induction medium with different formulations and conditions were as follows:
[0061] 1#: Stem Cell Basal Medium (Ste...
Embodiment 25a
[0067] Example 2.5a - Reductase Inhibition Experiment
[0068] Androgenetic alopecia (AGA) is due to the conversion of testosterone (T) into more active dihydrotestosterone (DHT) under the action of 5a-reductase. Hair follicles are sensitive to DHT. A large amount of DHT affects the development of hair follicles, leading to hair loss . Therefore, to judge whether a certain substance has hair growth function, whether it can inhibit 5a-reductase can be used as one of the judgment basis. The positive control drug in this embodiment is the hair loss treatment drug finasteride, which is a recognized drug inhibitor of 5a-reductase. The content of 5a-reductase is measured by its mRNA expression, and the detection process is as follows:
[0069] Human dermal papilla cells were cultured for 48 hours in the culture medium added with the aforementioned 4 kinds of umbilical cord mesenchymal stem cell-secreted factors obtained under different conditions, the medium added with finasteride...
Embodiment 3
[0087] Example 3. Experiments on Promoting Hair Follicle Culture in Vitro
[0088] The vibrissa follicles of juvenile mice can survive for a certain period of time in a specific medium in vitro. Hair can continue to grow while the follicle is alive. Different formulas contain different components in the cultured genes, and the survival time of hair follicles can be kept differently. The influence of different formulas of culture media on hair follicles can be judged according to the different growth lengths of hair.
[0089] After the hair follicle tissue was isolated from the upper lip of the mouse, the hair follicles were placed in different wells of a 24-well plate. Hair follicles were incubated for 14 days at 37°C, 5% carbon dioxide. Remove the ordinary culture medium, add the four kinds of polypeptide factor compositions obtained in Example 1 or the culture composition of the control group to the wells, measure the following indicators, and change the medium every three...
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