Non-feeder cell cultivation system culture medium for human embryonic stem cell and cultivation method therefor
An embryonic stem cell and feeder-free technology is applied in the field of stem cell culture to achieve the effect of low culture cost
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[0013] Methods and steps:
[0014] 1. Human embryonic stem cell culture medium formula (NB medium) without feeder cells and exogenous cytokines;
[0015] Utilize DMEM / F12 (11330057, DMEM NUTRIENT MIX F12, Gibco) N2 SUPPLEMENT (17502048, 100×, Gibco), B27 SUPPLEMENT (17504044, 50×, Gibco), β-ME (β-mercaptoethanol, Gibco) configuration provided by the present invention The medium: DMEM / F12, 1×N2, 1×B27, 0.1mM β-ME.
[0016] The components of each 50ML are as follows: DMEM / F12 48.5ml
[0017] N2 0.5ml
[0018] B27 1ml
[0019] β-ME 91μl
[0020] 2. Petri dish pretreatment
[0021] 1) Thaw the imported fetal bovine serum at 4°C, then divide it into 2ml / EP tubes and store at -20°C for later use;
[0022] 2) Thaw the serum in the EP tube at room temperature;
[0023] 3) Coat imported FBS to treat a 1-well plate (0.5ml) or a 6-well plate (1.5ml) at 4°C overnight or at least 4 hours at room temperature;
[0024] 4) Aspirate the FBS before use a...
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