Cells cultured with ReproFF2 retain the key characteristics of pluripotent cells

Maintanance of undifferentiated human iPS cells with ReproFF2

human iPS cell line 201B7
Feeder-free culture with ReproFF2: Passage 10, day 7
Feeder-dependent culture on MEF: Passage 48, day 3

1. Morphology and ALP activity

Human iPS cell line 201B7 (established in Kyoto University) was kept undifferentiated in feeder-free culture with ReproFF2, as confirmed by morphology (A) and alkaline phosphatase staining (B). These results looked comparable to those of feeder-dependently cultured iPS cells (C and D).

2. Expression of pluripotency markers

Human iPS cells cultured with ReproFF2 retained expression of pluripotency marker genes Oct 3/4, SSEA-4, TRA-1-60 and Nanog, similarly to cells cultured on feeder cells.

3. FACS analysis of pluripotency marker expression in iPS cells

Human iPS cells cultured with ReproFF2 showed profiles comparable to those of feeder-dependently cultured cells when analysed with FACS for pluripotency marker genes SSEA-4, TRA-1-60 and TRA-1-81.

4. Karyotype analysis of human iPS cells after long term culture with ReproFF2

Human iPS cells cultured for 30 passages using ReproFF2 showed no abnormality in karyotype analysis. This result shows ReproFF2 is suitable for long term culture of human iPS cells.

Maintanance of undifferentiated human ES cells with ReproFF2

human ES cell line KhES-1

Human ES cell line KhES-1 (established in Kyoto University) remained undifferentiated in feeder-free culture with ReproFF2, as confirmed by morphology (A), alkaline phosphatase staining (B) and Oct 3/4 expression (C). In collaboration with Kyoto University.