Mixed cell preparation and preparation method and application thereof
A cell preparation and cell technology, applied in the field of immune cells, can solve the problems of limited source of islets, inability to completely solve islet β cells, and high cost, so as to reduce or eliminate attacks on islet β cells, alleviate or treat type 1 diabetes, Promotes regenerative and repairing effects
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[0042] The present invention also provides a method for preparing the above-mentioned mixed cell preparation, the preparation method includes the following steps:
[0043] S1. Obtain the above-mentioned mesenchymal stem cells;
[0044] S2. Acquiring the above-mentioned regulatory T cells;
[0045] S3. The above solution is provided, and mesenchymal stem cells and regulatory T cells are added to the solution.
[0046] The preparation method of the mixed cell preparation of the present invention is simple and efficient, and the preparation can be completed only by blending mesenchymal stem cells, regulatory T cells and an appropriate amount of solution. It should be noted that the mesenchymal stem cells can be obtained by ex vivo tissue culture, or by purchasing commercially available mesenchymal stem cells; the regulatory T cells can be obtained by separating peripheral blood, or by purchasing commercially available Regulatory T cells are obtained.
[0047] Further, this step ...
Embodiment 1
[0056] 1. Obtain human umbilical cord mesenchymal stem cells
[0057] (1) Isolation of primary umbilical cord mesenchymal stem cells
[0058] After the human umbilical cord was removed from the operating table, it was immersed in physiological saline containing antibiotics and stored at 4°C;
[0059] Take out the human umbilical cord in the ultra-clean bench, rinse the residual blood in the umbilical artery and umbilical vein with D-PBS buffer solution, use hemostatic forceps and scissors to remove the washed arteries and veins, and then cut the human umbilical cord into 1mm 3 After the size of the tissue block, put it into a 200ml blue cap reagent bottle;
[0060] Add 30ml of type II collagenase with a mass volume ratio of 0.1% to the blue cap reagent bottle, place the blue cap reagent bottle in a constant temperature shaker for 6 hours, and then filter and collect primary umbilical cord mesenchymal stem cells with a 100-mesh sieve ;
[0061] (2), culture of primary umbil...
Embodiment 2
[0084] 1. Obtain human umbilical cord mesenchymal stem cells
[0085] Please refer to Example 1, which is the same as Example 1.
[0086] 2. Obtain regulatory T cells
[0087] Please refer to Example 1, which is the same as Example 1.
[0088] 3. Preparation of mixed cell preparations
[0089] Provide human umbilical cord mesenchymal stem cells 8×10 6 , CD4 + CD25 + Treg cells 1×10 7 One, and solution 20ml, wherein, this solution comprises the glucose that mass fraction is 5%, the NaCl that mass fraction is 0.45%, the human serum albumin that mass fraction is 0.5% and the water that mass fraction is 94.05%; Mesenchymal stem cells, CD4 + CD25 + Treg cells are mixed with the solution to prepare a fresh non-refrigerated cell suspension. In this mixed cell preparation, the concentration of human umbilical cord mesenchymal stem cells was 4×10 5 cells / ml, CD4 + CD25 + The concentration of Treg cells was 0.5×10 6 pieces / ml.
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