TeSR™-E8™

Protocol tips

Upstream tips	Protocol tips	Downstream tips
cells infected with Tet-on lentivirus	E8 medium supplemented with Y27632 (5 µM, Tocris) at 3.3 × 104 cells/cm2. The following day, E8 media was replaced with E6 media and cells were cultured for 2 days supplemented with CHIR99021 (10 µM, Selleck Chemical), after which CHIR99021 was removed and E6 media supplemented with 1 µg/mL Dox (Sigma) for 18 days until GFP+ induced myogenic progenitor cells (iMPCs) were sorted by FACS as described below. During differentiation of iMPCs, 10 ng/mL bFGF (R&D) was added starting at day 5 to enhance proliferation of GFP+ cells.

Upstream tips
cells infected with Tet-on lentivirus
Protocol tips
E8 medium supplemented with Y27632 (5 µM, Tocris) at 3.3 × 104 cells/cm2. The following day, E8 media was replaced with E6 media and cells were cultured for 2 days supplemented with CHIR99021 (10 µM, Selleck Chemical), after which CHIR99021 was removed and E6 media supplemented with 1 µg/mL Dox (Sigma) for 18 days until GFP+ induced myogenic progenitor cells (iMPCs) were sorted by FACS as described below. During differentiation of iMPCs, 10 ng/mL bFGF (R&D) was added starting at day 5 to enhance proliferation of GFP+ cells.

Publication protocol

E8 medium supplemented with Y27632 (5 µM, Tocris) at 3.3 × 104 cells/cm2. The following day, E8 media was replaced with E6 media and cells were cultured for 2 days supplemented with CHIR99021 (10 µM, Selleck Chemical), after which CHIR99021 was removed and E6 media supplemented with 1 µg/mL Dox (Sigma) for 18 days until GFP+ induced myogenic progenitor cells (iMPCs) were sorted by FACS as described below. During differentiation of iMPCs, 10 ng/mL bFGF (R&D) was added starting at day 5 to enhance proliferation of GFP+ cells.

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Manufacturer protocol

Download the product protocol from STEMCELL technologies for TeSR™-E8™ below.

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