Figure 4.

Comparison between regular passaging and cell recovery after cryopreservation. Six wells of H1 ESCs were dissociated by EDTA/PBS, harvested in E8 medium, and split into 24 equal portions. The cells were treated in triplicates as the table indicated. 1, Cells were directly plated into E8; 4. Cells were directly plated with ROCK inhibitor. 2 and 3, Cells were cryopreserved in E8 medium with 10% DMSO without (2) or with (3) ROCK inhibitor, and later recovered in E8 medium without ROCK inhibitor. 5 and 6, Cells were cryopreserved in E8 medium with 10% DMSO without (5) or with (6) ROCK inhibitor, and later recovered in E8 medium with ROCK inhibitor. A. Images of cells on matrigel-coated plate after different treatments. APS Staining was performed 48 hours after plating. B. Cryopreservation efficiency for cells recovered without ROCK inhibitor in E8 medium (Conditions 1, 2 and 3). The cell survival was analyzed 24 hours after plating, and normalized by the data obtained in Condition 1. C. Cryopreservation efficiency for cells recovered with ROCK inhibitor in E8 medium (Conditions 4, 5 and 6). The cell survival was analyzed 24 hours after plating, and normalized by the data obtained in Condition 4.