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Nano-structured films with super hydrophobic surfaces were fabricated by microwave-plasma enhanced chemical vapor deposition (MPECVD) method. It was found that water droplets with a volume from 10 to 30 μl rolled and bounced without wetting or spreading on the surfaces of the nanostructured films (nano films). After 3 days of cultivation, aggregation of embryoid body (EB) cells from embryonic stem (ES) cells were formed as microspheres on nano films coating in petri disks. Growth of ES cells was shown to be the best on the nano films with a 150 o water contact angle. The EBs derived from ES cells cultured in droplets on the nano films grew over the cultivation time until the 7 th day on which EBs began to spread out. The results showed that the nano-structured film with a super-hydrophobic surface is a useful material for obtaining EBs from undifferentiated ES cells, this is anticipated to be a stem cell source for transplantation due to their pluripotency and indefinite proliferation.