Large-scale preparation method of mesenchymal stem cell factors
A technology for large-scale preparation of mesenchymal stem cells, which is applied in the field of large-scale preparation of mesenchymal stem cell factors, can solve the problems of small amount of mesenchymal stem cell factors, difficulty in ensuring product uniformity, and unscalable separation and purification process, and achieve the goal of preparing The process conditions are stable and reasonable, and the product quality stability is realized, which is conducive to the effect of stability
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Embodiment 1
[0043] A large-scale preparation method of mesenchymal stem cell factors, comprising the following steps:
[0044] 1. Select mesenchymal stem cells, after in vitro primary culture and subculture to 4th generation, digest and count the 4th generation cells, and transfer them to the cell factory for expansion culture;
[0045] 2. When the expanded and cultured cells grow to a confluence of 70-85%, discard the original medium, add the induction medium to the cell factory, continue the culture, and collect the induction medium;
[0046] 3. Circulate and filter the collected induction medium through the hollow fiber membrane for 5 times, collect the first crude pure liquid, pump the first crude pure liquid to the tangential flow ultrafiltration membrane bag, and circulate and filter to the first crude pure liquid Concentrate the volume to 1 / 50 of the original volume, then add PBS buffer to the concentrated crude pure solution to make up to the original volume to obtain the second c...
Embodiment 2
[0049] A large-scale preparation method of mesenchymal stem cell factors, comprising the following steps:
[0050] 1. Select mesenchymal stem cells, after in vitro primary culture and subculture to 4th generation, digest and count the 4th generation cells, and transfer them to the cell factory for expansion culture;
[0051] 2. When the expanded and cultured cells grow to a confluence of 70-85%, discard the original medium, add the induction medium to the cell factory, continue the culture, and collect the induction medium;
[0052] 3. Circulate and filter the collected induction medium through the hollow fiber filter membrane for 8 times, collect the first crude pure liquid, pump the first crude pure liquid to the tangential flow ultrafiltration membrane bag, and circulate and filter to the first crude pure liquid Concentrate the volume to 1 / 30 of the original volume, then add PBS buffer to the concentrated crude pure solution to make up to the original volume to obtain the s...
Embodiment 3
[0055] A large-scale preparation method of mesenchymal stem cell factors, comprising the following steps:
[0056] 1. Select mesenchymal stem cells, after in vitro primary culture and subculture to 4th generation, digest and count the 4th generation cells, and transfer them to the cell factory for expansion culture;
[0057] 2. When the cells grow to a confluence of 70-85%, discard the original medium, and add induction medium according to half the volume of the original medium, and continue to cultivate and adjust the parameters: 37 ° C, 5% CO 2 , 5% O 2 After 24 hours, adjust the culture parameters to: 37°C, 5% CO 2 , 20%O 2 After 48 hours, half the amount of induction medium was collected, and an equal amount of fresh induction medium was added to the cell factory, and the parameters were adjusted to 37°C, 5% CO 2 , 5% O 2 , continue to cultivate; collect all induction medium after 72h;
[0058] 3. Pass the collected induction medium through a 0.1um hollow fiber filter...
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