Preparation method and application of polymer coating for long-term culture of human adipose stem cells in vitro
A technology of polymer coating and adipose stem cells, which is applied in the direction of cell culture support/coating, cell culture active agent, biochemical equipment and methods, etc., can solve the problem of not being able to ensure the pluripotency of stem cells, reducing the ability of differentiation, Pathogen transfer infection and other problems, to achieve good optical transparency, good biocompatibility, and promote the effect of proliferation
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Embodiment 1
[0032] Weigh 0.1g methacryloxyethyltrimethylammonium chloride solution (E), 0.1g methacrylate-2-(diethylamino)ethyl (K), 0.1g cyclohexyl methacrylate ( L) Add 0.3g of 1-methyl-2-pyrrolidone, add 0.015g of 1-hydroxycyclohexyl benzophenone and 0.03g of N,N'-methylenebisacrylamide to form a polymer solution, and use a pipette to absorb the polymer The substance solution was added dropwise on the PET film, and then the cover glass was covered on the droplet, and the air was squeezed out gently. After irradiating with 365nm ultraviolet light for 30min, the PET film was peeled off, and the glass slide was placed in an oven at 40°C for 12h. Finally, it was rinsed twice with acetone and ethanol, and air-dried to obtain the polymer coating E1K1L1.
[0033] Human adipose-derived stem cells were placed at a density of 1×10 5 / mL seeded on the polymer coating for long-term culture. Collect the second generation human adipose stem cells, adjust the cell concentration to 1×10 5 / mL, inoc...
Embodiment 2
[0038] Weigh 0.15g methacryloxyethyltrimethylammonium chloride solution (E), 0.05g methacrylate-2-(diethylamino)ethyl (K), 0.1g cyclohexyl methacrylate ( L) Add 0.3g of 1-methyl-2-pyrrolidone, add 0.015g of 1-hydroxycyclohexyl benzophenone and 0.03g of N,N'-methylenebisacrylamide to form a polymer solution, and use a pipette to absorb the polymer The substance solution was added dropwise on the PET film, and then the cover glass was covered on the droplet, and the air was squeezed out gently. After irradiating with 365nm ultraviolet light for 30min, the PET film was peeled off, and the glass slide was placed in an oven at 40°C for 12h. Finally, it was rinsed twice with acetone and ethanol, and air-dried to obtain the polymer coating E3K1L2.
[0039] Human adipose stem cells were cultured and induced, stained and characterized according to the steps in Example 1.
Embodiment 3
[0041] Weigh 0.15g methacryloxyethyltrimethylammonium chloride solution (E), 0.05g hydroxyethyl methacrylate (A), 0.1g cyclohexyl methacrylate (L) and add 0.3g 1- Add 0.015g of 1-hydroxycyclohexyl benzophenone and 0.03g of N,N'-methylenebisacrylamide to methyl-2-pyrrolidone to form a polymer solution, use a pipette to absorb the polymer solution and drop it on the PET film Place a coverslip over the droplet and squeeze gently to expel the air. After irradiating with 365nm ultraviolet light for 30min, the PET film was peeled off, and the glass slide was placed in an oven at 40°C for 12h. Finally, it was rinsed twice with acetone and ethanol, and air-dried to obtain the polymer coating E3A1L2.
[0042] Human adipose stem cells were cultured and induced, stained and characterized according to the steps in Example 1.
[0043] figure 1 It is the infrared spectrogram of the polymer layer E3K1L2 of embodiment 2, and the mixed monomer C=C double bond is at 1630cm before photopolymer...
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