Method for three-dimensional culture of fibrous ring derived stem cells based on Collagel gel bracket method
A technology of three-dimensional culture and annulus fibrosus, applied in the fields of biotechnology and cell culture, can solve the problems of unsatisfactory effect of 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
[0044] The Collagel culture medium matrix described in the present embodiment comprises following composition:
[0045] Reagent A: 5×MEM, 288 μl;
[0046] Reagent B: FBS, 144 μl;
[0047] Reagent C: Hepes (1M), 36 μl;
[0048] Reagent D: 0.1M NaOH and 5×MEM mixture, 72μl;
[0049] Collagel: 3mg / ml COL1 mixed with 0.01M HCl, 1260μl.
[0050] 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 mixed with 0.01M HCl solution) and 72 μl of reagent D (mixed solution of 0.1M NaOH and 5×MEM) 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 the obtained Need Collagel culture substrate, 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.
[0051] Se...
Embodiment 2
[0056] The Collagel culture medium matrix described in the present embodiment comprises following composition:
[0057] Reagent A: 5×MEM, 288 μl;
[0058] Reagent B: FBS, 144 μl;
[0059] Reagent C: Hepes (1M), 36 μl;
[0060] Reagent D: 0.1M NaOH and 5×MEM mixture, 72μl;
[0061] Collagel: 3mg / ml COL1 mixed with 0.01M HCl, 1260μl.
[0062] 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 mixed with 0.01M HCl solution) and 72 μl of reagent D (mixed solution of 0.1M NaOH and 5×MEM) 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, Obtain the required Collagel culture substrate for later use. It should be noted that the storage and operation of all the above reagents need to be kept at 4°C or on ice.
[0063] Selec...
Embodiment 3
[0067] The Collagel culture medium matrix described in the present embodiment comprises following composition:
[0068] Reagent A: 5×MEM, 288 μl;
[0069] Reagent B: FBS, 144 μl;
[0070] Reagent C: Hepes (1M), 36 μl;
[0071] Reagent D: 0.1M NaOH and 5×MEM mixture, 72μl;
[0072] Collagel: 3mg / ml COL1 mixed with 0.01M HCl, 1260μl.
[0073] 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 mixed with 0.01M HCl solution) and 72 μl of reagent D (mixed solution of 0.1M NaOH and 5×MEM) 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, Obtain the required Collagel culture substrate for later use. It should be noted that the storage and operation of all the above reagents need to be kept at 4°C or on ice.
[0074] Selec...
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