Prof

Kanji Miyamoto

e-mail

kmiyamot@cc.tmit.ac.jp

Development of nuclear transplantation techniques using somatic cells for the production of cloned ES cells

The creation of cloned ES cells by nuclear transplantation using somatic cells in primates including human beings has never been achieved, although it has been successfully performed in mice. In the future, it will be important to establish methods for producing ES cells that do not trigger rejection via transplantation of nuclei from somatic cells derived from a patient to enucleated ova in order to therapeutically transplant the differentiated cells targeted in the patient. We are aiming to produce monkey ES cells via nuclear transplantation (ntES) using somatic cells after enucleating an ovum of a monkey as a preliminary step toward establishing nuclear transfer-derived human ES cell lines because of the ethical issues involved in human experiments. The final goal is to establish nuclear transfer human ES cell lines (ntES)for individual patients themselves.

Development of induction methods for differentiation from nuclear transfer mouse ES cells to various types of cells

Self-beating myocardial cells were induced by cultivating mouse ES cells established from mouse fertilized ova to produce embryoid bodies. In addition, the molecular biological characteristics of the myocardial cells were examined. Differentiation induction is currently performed on various cells using mouse ES cells by nuclear transplantation using somatic cells (nuclear transfer ES :ntES). Specifically, nerve cells such as cells producing dopamine which are neurotransmitters necessary for the treatment of Parkinson's disease, hematopoietic stem cells which are used in the treatment of leukemia, islet cells which secrete insulin required for the treatment of diabetes, retinal cells, cartilage cells, myocardial cells and skeletal muscle cells are being investigated. Furthermore, the effectiveness of cell transplantation is being examined by transplanting differentiated cells to disease animal models.

Establishment of methods for the proliferation of somatic stem cells and embryonic stem cells

Bone marrow transplantation uses marrow cells for treating diseases such as leukemia. Moreover, a treatment has been developed in which stem cells isolated from bone marrow were transplanted to patients with cardiac infarction to regenerate blood vessels. More recently, cell transplantation has been performed by isolating hematopoietic stem cells from blood derived from the umbilical cord for transplantation. However, there are various problems. Stem cells cannot be isolated in sufficient quantity for transplantation to all affected patients, and so in vitro proliferation of stem cells is required. We have investigated a combination of cytokines to develop a system for the proliferation of stem cells and evaluated the use of human amnion cells as feeder cells. Recently, ES-like cells have been confirmed from umbilical cord blood. It is important to develop a system for isolating and proliferating these cells. In addition, it is also important to develop a safe culture system for proliferating ES cells aiming at clinical application. To accomplish these goals, we have developed human-amnion cells as feeder cells. In future, it will be necessary to develop a safe cell-free system.

Recent Publications

1. Miyamoto. K., Hayashi. K., Suzuki. T., Ichihara. S., Yamada. T., Kano. Y., Yamabe. T. and Ito. Y. (2004) Human placenta feeder layers support undifferentiated growth of Primate embryonic stem cells. Stem Cells. 22. 433-440.
2. Ito. Y., Konno. T and Miyamoto. K. (2004) Culture of embryonic stem cells. Bio Industry. 21, 5-11.
3. Ito. Y., Nogawa. M., Konno. T., Makino. H., Hasuda. H., Wu. Z., Miyamoto. K. and Ikebuchi. K. (2004) Biomaterials for in vitro expansion of stem cells. Regenerative Medicine, 3, 55-61.
4. Kano. Y., Nohno. T., Hasegawa. T., Takahashi. R., Hiragami. F., Kawamura. K., Ieama. M.K., Motaoda. H and Miyamoto. K. (2002) Immunosuppressant FK506 induces neurite outgrowth in PC12 mutant cells with impaired NGF promoted neuritogenesis via a novel MPA kinase signaling pathway. Neurochemical Research, 12, 1655-1661.
5. Kano. Y., Takaguchi. S., Nohno. T., Hiragami. F., Iwama. M., Miyamoto. K and Takehara. M. (2002) Chinese medicine induces neurite outgrowth in PC12 mutant cells incapable of differentiating. American Journal of Chinese Medicine, 30, 287-295.
6. Kano. Y., Hiragahi. F., Kawamura. K., Takaguchi. S., Kimata. Y., Poffenberger. C.K., Iwama. M., Yoshimizu. A., Nishimura. S., Miyamoto. K and Namba.M. �i2002�jEsatablishment of a drug-hypersensitive PC12 mutant clone deficient in nerve growth factor induced neurine outgrowth. Tiss. Cult. Res. Commun. 20, 145-153.
7. Katayama. Y., Miyamoto. K., Takenaka. K., Imajyo. K., Shigenaga. K. and Harada.M.,(2001) Chromosome analysis after ex vivo expansion of CD34(+) cells from human cord blood. Cancer Genet Cytogen. 125, 161-162.
8. Hayashi. K., Chen. H.L., Hayashi. H., Koirala. T.R., Ohara. N., Teramoto. N., Oka. T., Yoshio. T., Takahashi. K., Miyamoto. K., Fujimoto., Yoshikawa. T and T. Akagi. (2001) Cyno-EBV(EBV-related herpvirus from cynomolgus macaques) induces rabbit malignant lymphomas and their tumor cell lines frequently show specific chromosomalities. Lab Inves. 79, 823-835.