Method for inducing pancreas islet cells differentiated by stem cells in vitro to form pancreas islet-like structure
A technology for stem cell differentiation and islet cells, applied in cell dissociation methods, artificial cell constructs, biochemical equipment and methods, etc., can solve problems such as low induction rate and negative impact on product quality, and achieve high induction rate and application effect Good, the effect of high product cell quality
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Embodiment 1
[0030] Example 1 In vitro induction of stem cell-differentiated islet cells to form islet-like structures:
[0031] S1. Obtaining mesenchymal stem cells by separating and culturing from raw material tissues;
[0032] S2, subculture of the obtained mesenchymal stem cells;
[0033] S3. Inducing and differentiating the obtained passaged mesenchymal stem cells into islet-like cells.
[0034] The raw material tissue in S1 includes human placenta tissue.
[0035] The isolation culture described in S1 includes: washing the raw material tissue, cutting it into pieces, digesting it with digestive enzymes, centrifuging to obtain a cell pellet, and inoculating it in a cell culture medium after washing.
[0036] The above-mentioned digestive enzymes include type I collagenase, and the digestion time is 45 minutes.
[0037] For the above centrifugation, the condition is to centrifuge at 1200 rpm for 5 minutes; for the above washing, use PBS to wash 3 times.
[0038] After being inocula...
Embodiment 2
[0049] Example 2 In vitro induction of stem cell-differentiated islet cells to form islet-like structures:
[0050] S1. Obtaining mesenchymal stem cells by separating and culturing from raw material tissues;
[0051]S2, subculture of the obtained mesenchymal stem cells;
[0052] S3. Inducing and differentiating the obtained passaged mesenchymal stem cells into islet-like cells.
[0053] The raw material tissue in S1 includes adipose tissue.
[0054] The isolation culture described in S1 includes: washing the raw material tissue, cutting it into pieces, digesting it with digestive enzymes, centrifuging to obtain a cell pellet, and inoculating it in a cell culture medium after washing.
[0055] The above-mentioned digestive enzymes include type II collagenase, and the digestion time is 60 minutes.
[0056] For the above centrifugation, the condition is to centrifuge at 1500 rpm for 8 minutes; for the above washing, use PBS to wash 5 times.
[0057] After being inoculated in ...
Embodiment 3
[0068] Example 3 In vitro induction of stem cell-differentiated islet cells to form islet-like structures:
[0069] S1. Obtaining mesenchymal stem cells by separating and culturing from raw material tissues;
[0070] S2, subculture of the obtained mesenchymal stem cells;
[0071] S3. Inducing and differentiating the obtained passaged mesenchymal stem cells into islet-like cells.
[0072] The raw material tissue in S1 includes placental tissue.
[0073] The isolation culture described in S1 includes: washing the raw material tissue, cutting it into pieces, digesting it with digestive enzymes, centrifuging to obtain a cell pellet, and inoculating it in a cell culture medium after washing.
[0074] The above-mentioned digestive enzymes include type I collagenase or type II collagenase, and the digestion time is 50 minutes.
[0075] For the above centrifugation, the condition is 1400 rpm for 6 minutes; for the above washing, PBS is used for washing 4 times.
[0076] After bein...
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