Method for acquiring monkey haploid neural stem cells
A neural stem cell, haploid technology, applied in the field of bioengineering
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[0026] 1. Preparation of a pluripotent monkey embryonic stem cell culture medium
[0027] After improvement, the culture medium for the growth of haploid monkey stem cells contains D / F12 medium with 20% KOSR and 0.1mM non-essential amino acids, and other supplements in the medium are 0.1mM β-mercaptoethanol, 100U / ml penicillin, 0.1 mg / ml streptomycin, 2mM L-Glutamine, 5ng / ml fibroblast growth factor (bFGF), 10ng / ml human leukemia inhibitory factor (LIF), 0.5μM PD0325901, 3μM CHIR99021, 10μM SB203580 and 10 μM of SP600125.
[0028] 2. Obtaining haploid neural stem cells from rhesus monkeys
[0029] The macaque haploid embryonic stem cells in culture were digested into single cells with 0.05% trypsin, resuspended with EB culture medium and cultured in a suspension culture dish. On the first day of differentiation, 10 μM Y-27632 was added to the EB culture medium to reduce cell apoptosis. On the 5th day of differentiation, the EB culture medium was replaced with neural inducti...
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