Method for three-dimensionally culturing bone marrow mesenchymal stem cells based on Collagel scaffold method
A bone marrow mesenchymal, three-dimensional culture technology, applied in the field of biotechnology and cell culture, can solve the problems of unsatisfactory three-dimensional cell culture technology, and achieve the effect of ensuring tissue moisture, enhancing adhesion ability, and not easy to fall off
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Embodiment 1
[0043] The Collagel culture medium matrix described in the present embodiment comprises following composition:
[0044] Reagent A: 5×MEM, 288 μl;
[0045] Reagent B: FBS, 144 μl;
[0046] Reagent C: Hepes (1M), 36 μl;
[0047] Reagent D: 0.1M NaOH and 5×MEM mixture, 72μl;
[0048] Collagel: 3mg / ml COL1 mixed with 0.01M HCl, 1260μl.
[0049] Take 288 μl of Reagent A (5×MEM), 144 μl of Reagent B (FBS) and 36 μl of Reagent C (Hepes, 1M) in Test Tube 1 for thorough mixing and set aside; another 1260 μl of Collagel (3mg / ml COL1 with 0.01M HCl mixed solution) and 72 μl of reagent D (0.1M NaOH and 5×MEM mixed solution) were placed in test tube 2 for thorough mixing, and the pH of the mixed solution was adjusted to 7.0 for later use; the mixed solutions in test tube 1 and test tube 2 were mixed to obtain Required Collagel culture medium, spare. It should be noted that the storage and operation of all the above reagents need to be kept at 4°C or on ice.
[0050] Select bone marro...
Embodiment 2
[0055] The Collagel culture medium matrix described in the present embodiment comprises following composition:
[0056] Reagent A: 5×MEM, 288 μl;
[0057] Reagent B: FBS, 144 μl;
[0058] Reagent C: Hepes (1M), 36 μl;
[0059] Reagent D: 0.1M NaOH and 5×MEM mixture, 72μl;
[0060] Collagel: 3mg / ml COL1 mixed with 0.01M HCl, 1260μl.
[0061] Take 288 μl of Reagent A (5×MEM), 144 μl of Reagent B (FBS) and 36 μl of Reagent C (Hepes, 1M) in Test Tube 1 for thorough mixing and set aside; another 1260 μl of Collagel (3mg / ml COL1 with 0.01M HCl mixed solution) and 72 μl reagent D (0.1M NaOH and 5×MEM mixed solution) were placed in test tube 2 for thorough mixing, and the pH of the mixed solution was adjusted to 7.0-7.2 for later use; mix the mixed solutions in test tube 1 and test tube 2 , to obtain the required Collagel culture substrate, and set aside. It should be noted that the storage and operation of all the above reagents need to be kept at 4°C or on ice.
[0062] Select...
Embodiment 3
[0066] The Collagel culture medium matrix described in the present embodiment comprises following composition:
[0067] Reagent A: 5×MEM, 288 μl;
[0068] Reagent B: FBS, 144 μl;
[0069] Reagent C: Hepes (1M), 36 μl;
[0070] Reagent D: 0.1M NaOH and 5×MEM mixture, 72μl;
[0071] Collagel: 3mg / ml COL1 mixed with 0.01M HCl, 1260μl.
[0072] Take 288 μl of Reagent A (5×MEM), 144 μl of Reagent B (FBS) and 36 μl of Reagent C (Hepes, 1M) in Test Tube 1 for thorough mixing and set aside; another 1260 μl of Collagel (3mg / ml COL1 with 0.01M HCl mixed solution) and 72 μl reagent D (0.1M NaOH and 5×MEM mixed solution) were placed in test tube 2 for thorough mixing, and the pH of the mixed solution was adjusted to 7.0-7.2 for later use; mix the mixed solutions in test tube 1 and test tube 2 , to obtain the required Collagel culture substrate, and set aside. It should be noted that the storage and operation of all the above reagents need to be kept at 4°C or on ice.
[0073] Select...
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