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Life Technologies Parkinson’s disease cell models

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A BC D E FG H Figure 2. iPSC generation from PD-3 donor (A–D) or Ctrl-1 individual (E–H) fibroblasts. iPSC generation was performed according to the User Guide for the CytoTune®-iPS Sendai Reprogramming Kit. Briefly, fibroblasts were cultured in DMEM with 20% FBS for 2 days on a 6-well plate coated with Attachment Factor (A, E). Cells were transduced overnight with the CytoTune®-iPS Sendai Reprogramming Kit at an MOI of 3. At day 7 post-transduction (B, F) cells were harvested, counted using the Countess® Automated Cell Counter, and plated onto 10 cm MEF culture dishes at 0.5, 1, 2, or 3 x 105 cells/dish. Colonies were picked between days 20 and 26. Representative colonies before picking are shown in C on day 22 or in G on day 26. Picked colonies were transferred to a 12-well MEF culture plate. Individual colonies were further expanded (D, H), cryopreserved, and characterized. Instrument highlight—EVOS® XL Imaging System The cells were monitored and imaged throughout the reprogramming process using the EVOS® XL Imaging System, a transmitted light microscope that combines a digital camera, LCD display, and USB device storage. iPSC colonies were picked and passaged into new dishes between 20 and 26 days after the CytoTune® reagents were added to the fibroblasts. 12 Life Technologies | Parkinson's cell model

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