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Embryonic stem (ES) cells have been known to differentiate into various progenitor cells. In this study, we investigated the differentiation capacity of mouse ES cells into pancreatic hormone-secreting cells, and insulin-secreting cells. ES cells were cultured in Dulbecco's modified Eagle medium (DMEM) after removal of leukemia inhibitory factor (LIF) for 3 days and then transferred in DMEM supplemented with retinoic acid or activin A. When the culture of embryoid bodies (EBs) derived from ES cells in DMEM added with activin A (2 × 10-9 M or 2 × 10-10 M) for 5 days was performed, endoderm marker genes, GATA4, Sox17 and Foxa2 were expressed in the EBs. However, at 6 days of culture, expression of Sox17 was not observed. When EBs were cultured with activin A (2 × 10-9 M) for 6 days, and followed by 6 days of culture with retinoic acid (10-6 M), expression of the pancreatic cell marker genes, GATA4 and Fxa2, were continued, and pancreatic islet genes, insulins 1 and 2, glucagons and somatostatin, w...

In nuclear transplantation, especially in enucleation, rotating the oocyte to a certain position has been an important work. Therefore, we developed an oocyte rotation system by using electrostatic force, which is known to be excellent in handling minute objects. This system has become the basic technology for the automated nuclear transplantation system. Mouse oocyles were rotated at a speed of about 60 deg/s by electrostatic phenomena (i.e. electrorotation and dielectrophoresis), and the direction of rotation could be reversed. In order to confirm the electrostatic effect on the fertilization and developmental ability of oocytes, we performed IVF and nuclear transplantation with rotated oocytes. The result was that the fertilization and developmental ability of the rotated oocytes was the same as that of the control, proving that rotation by electrostatic force does not influence the fertilization and development ability of oocytes.

A square case containing ova and culture medium was vibrated with a voice coil motor to concentrate the ova at the center of the case. Only those ova which conform to the memorized shape are then selected with a picture processing device and are positioned one by one at the point of a holding pipette. A system with the functions mentioned was experimentally constructed. This system can automatically position any ovum in a certain position, thereby shortening the time of the process to search out an ovum and position it at the point of the holding pipette.