Lentivirus packaging and infection
This protocol is extracted from research article:
Deficiency of TRPM2 leads to embryonic neurogenesis defects in hyperthermia
Sci Adv, Jan 1, 2020; DOI: 10.1126/sciadv.aay6350

The production of lentivirus was obtained by transfecting the core and packaging plasmids into 293FT cells using GenEscort I (Nanjing Wisegen Biotechnology). The virus was gathered at 24, 48, and 72 hours after changing the medium 6 hours after transfection. The primary NSCs for Western blot and immunofluorescence were seeded in 6- or 24-well plates, which were coated with laminin (Invitrogen) and poly-d-lysine (Sigma) (both 10 μg/ml) in advance. Twenty-four hours later, half of the medium was changed with proliferation medium without PS. Lentivirus was then added to each well and maintained for 8 hours. Meanwhile, to improve the infection efficiency, polybrene (2 μg/ml) was mixed into the medium. Forty-eight hours later, to induce a differentiation state, the medium was displaced with low-glucose DMEM (Gibco) supplemented with 1% FBS (Invitrogen), 1% PS, and 2% B27 (with vitamin A).

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