mESC Differentiation Images

Images for iMEF Feeder Cells:P13 iMEFS were received from Dr. Gul Afshan and, before propagating the flask on 4/25/2015

Figures 1-2 were taken.

Figure 1: P13 iMEFS (Flask A-9) dead cells at 20X 04/25/2015 Figure 2: P13 iMEFS (A-9X) at 10X 04/25/2015

One day after P14 iMEFS were propogated, reflective flakes were seen in all 10 flasks. Figures

3-4 show the flakes inside one of these flasks.

Figure 3: P14 iMEFs (Flask 4-1) at 10X 04/26/2015 Figure4: P14 iMEFs (Flask 4-2) at 10X 04/26/2015

Three days after P14 iMEFS were observed to contain flakes, the flasks were observed once

again. Figures 5-15 show the flakes inside one of these flasks along with the morphology,

confluency and some observed debris.

Figure 5: P14 iMEFs (Flask A-9.1X) at 10X 4/29/2015 Figure 6: P14 iMEFs (Flask A-9.1X) at 10X 4/29/2015





Figure 15: P14 iMEFs (Flask A-9.5X) at 10X 4/29/2015

The cells were observed again on 5/04/2015 prior to bleaching. The cells shown in Figure 16

were at about 90% confluency, but large amounts of cell debris and morphology were observed

in areas of the flask, therefore they were deemed too differentiated to continue propagating and

banking. These flasks are treated differently from those in Figure 17 since they were washed and

fed fresh media after flaking was observed. The cells in Figure 17 were ata low confluency and

show a similar beading morphology observed in Figure 11.

Figure 16: P14 iMEFs (Flask A-9.10X) at 10X 5/04/2015 Figure 17: P14 iMEFs (Flask A-9.8) at 10X 5/04/2015

Figures 18 and 19 show the results of Water Bears A-Taq’s feasibility experiment for the hanging drop method.

Figure 18: Image of upturned lid containing 20 uL droplets Figure 19: Image of TC dish with 20 uL droplets showing that none were lost

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mESC Differentiation Images