The indication for allogeneic stem cell transplantation (allo-SCT) have been expanded nowadays because many stem cell sources became available and new conditioning procedures such as reduced intensity stem cell transplantation (RIST) have been developed. Stem cell sources can be classified into bone marrow cells, peripheral blood stem cells, cord blood cells and every source derived from related or unrelated donors. Also, HLA mismatched transplantation has been studied especially in haploidentical donors. Now we must select the most compatible stem cell source for the recipient condition and disease status. RIST has expanded the indication of allo-SCT because of low regimen related toxicity. However, evaluation of graft versus leukemia (GVL) effect and control of graft versus host disease (GVHD) are still unresolved problems. Further investigations of the therapy of chronic GVHD and other posttransplant problems are warranted to improve the outcome and quality of life of the patients.

In the present study, the effects of liquid crystal-related compounds on the megakaryocytopoiesis and thrombopoiesis were evaluated in vitro using CD34+ cells prepared from human placental and umbilical cord blood (CB). About 20 kinds of compounds were tested for their effects on the clonal growth of CB CD34+ megakaryocytic progenitor cells (CFU-Meg) in plasma clot culture. The compounds, dissolved in DMSO, were added to the cultures within a concentration range of 10-100 nM. When used alone, none of the compounds supported the clonal growth of CFU-Meg. However, when thrombopoietin (TPO) was used as a growth factor, three compounds increased CFU-Meg clonal growth significantly, producing approximately 1.3-1.4 fold increases in the total number of megakaryocyte colonies in comparison with the control. These compounds promoted mainly mature CFU-Meg-derived small colonies, suggesting that their target is relatively mature CFU-Meg. These effective compounds were examined in liquid culture supplemented with TPO alone for 14 days. Although there was no evident promotion of the total number of cells harvested from the culture, two compounds suppressed cell growth significantly. Only one compound enhanced the generation of CFU-Meg in the harvested cells. Although these results do not indicate a strong correlation between the chemical structure of each compound and biological effectiveness, the incorporation of phenylpyridine and phenylpyrimidine and binding of a hydroxyl residue into the structure may play an important role in the activity. Thus, liquid crystal-related compounds whose biological action was previously unknown have been shown to act as regulators of hematopoiesis.

The discovery of the double-helical structure of DNA, the elucidation of the genetic code, and the determination of the three-dimensional structure of several proteins are some of the outstanding achievements of biochemistry and life sciences in the latter half of the last century. Proteins play key roles in almost all the biological processes and the biological function of a protein depends on its conformation which is defined as the three-dimensional arrangement of the atoms of a molecule. The three-dimensional structure, however, is not rigid but fluctuated. Structural fluctuation plays an important role in bio-macromolecules. How about "functional fluctuation" in biological systems? The present review proposes that functional fluctuation is also very important for understanding the mechanism of supramolecules, biological processes in living cells, and the interaction between biological systems. This new theme is pretty well supported by our recent experiments for neuro-immune crosstalk, gene transfection with cationic liposomes, and cell signaling in embryonic stem cells.

After the gut endoderm is formed, subsequent permissive induction signals from the notochord allows the pancreas to emerge and growout from a specific site of the embryonic gut epithelium. The first pancreas-specific gene, pdx1, is expressed around this stage. Interaction between pancreatic epithelium and the surrounding mesenchyme allows the pancreas bud to further grow and differentiate to form ductal, exocrine or endocrine lineages. Several lines of evidence from gene knockout mice and cell lineage studies suggest that a common pancreas stem cell first gives rise to exocrine and endocrine progenitor cells. The endocrine progenitor then give rise to four different endocrine cells: alpha, beta, delta, and PP cells, although it is not known how and when these cells arise. We have focused our studies on the understanding of endodermal induction and organogenesis of the pancreas. We specifically aimed at the isolation of pancreatic stem cells and the development of functional pancreatic endocrine beta cells in culture. For this aim, we used ES cells as a model system. Here, I review the literature on the development and regeneration of the pancreas. Some recent results on growing pancreatic cells from ES (embryonic stem) cells.

Regenerative medicine is a promising approach to treat patients with severe cardiac failure. Since embryonic stem cells (ES cells) easily differentiate into cardiomyocytes, ES cells are thought to be a good candidate resource for cardiac cell transplantation therapy. However, molecular mechanism of cardiac differentiation is still largely unknown. Here we discuss our present approach to understand the mechanism of cardiogenesis at the molecular level as well as novel genes and cascades that are important for cardiac differentiation. Further observation will help to establish the new strategy of regenerative medicine for patients with cardiac failure.

Bone morphogenetic protein (BMP) signaling is involved in astrocytic differentiation of neural precursor cells. In the nuclei, BMP-downstream transcription factors, i.e. Smad proteins, induce expression of astrocyte-specific genes in cooperation with another cytokine signaling, while inhibiting neuronal and oligodendrocytic differentiation-inducing transcription factors. This cross-inhibitory mechanism contributes to exclusive astrocytic differentiation. Crosstalk among cytokine signaling pathways thus is important for the development of the central nervous system.

Periodontal ligament (PDL) has a heterogeneous cell population, where some of the cells may be capable of differentiating into either cementoblasts or osteoblasts. Recently, C 2 H 2 zinc finger transcription factor Osterix has been reported. Osterix is one of the master regulators of bone cell differentiation and it has two different isoforms. According to a recent report, osteogenic differentiation of murine embryonic stem cells can be induced by overexpression of Osterix. The purpose of this study was to investigate about the expression of Osterix on human PDL (hPDL), and whether the osteogenic differentiation of hPDL cells can be induced by overexpression of Osterix. hPDL cells were obtained from healthy human teeth indicated for extraction for orthodontic treatment. All procedure used in this study was approved by the local ethical committee of Tokyo Medical and Dental University. To investigate expression of Osterix mRNA in hPDL tissues and cells, RT-PCR experiments were performed. Two different isoform Osterix expression vectors were made and transiently transfected into hPDL cells. Osteogenic differentiation was assessed by RT-PCR for genes associated with the osteoblast lineage such as Osteopontin, Osteocalcin, and Bone Sialoprotein. RT-PCR analyses showed that osterix mRNA was expressed in both hPDL tissue and cells. The expression of Osterix short isoform was higher than that of the long isoform. Overexpression of Osterix induced upregulated expression of Bone Sialoprotein mRNA. In expression levels of Osteopontin and Osteocalcin mRNA, compared to the control, no difference was observed. In conclusion, Osterix plays important roles in the osteoblastic differentiation in hPDL cells and modulates the mineralization.

Both normal somatic stem cells and cancer cells are thought to be capable of unlimited proliferation. Paradoxically, however, some cancers seem to contain stem-like cells (cancer stem cells). There is increasing evidence that cancers might contain their own stem cells. Many cancers, like normal organs, seem to be maintained by a hierarchical organization that includes slowly dividing stem cells,rapidly dividing transit amplifying cells (precursor cells), and differentiated cells. Malignant gliomas, for example,often contain both undifferentiated and differentiated cells and sometimes contain cells that express neuronal markers as well as cells that express glial markers, suggesting that they may contain multipotent neural stem cell-like cells. We have shown that some cancer cell lines contain a small side population (SP), which, in many normal tissues, is thought to contain the stem cells of the tissue. We provide evidence that SP cells in the C6 glioma cell line can produce both neurons and glial cells and thus have cancer stem cell property. Taken together with studies on normal neural stem cells, studies on cancer stem cells will help us to understand a link between normal stem cells and cancer stem cells.

Gene therapy for advanced breast cancer is anticipated to be a useful therapeutic approach. Strategies in ongoing clinical protocols can be divided into four groups: (1) suppression of oncogenes or transfer of tumor-suppressor genes: (2) enhancement of immunological response: (3) transfer of suicide genes: (4) protection of bone marrow using drug resistance genes. We have started a clinical study of multidrug resistance (MDR1) gene therapy. Patients received high dose chemotherapy and autologous peripheral blood stem cell transplantation (PBSCT) with MDR1-transduced hematopoietic cells, and then were treated with docetaxel. Three patients have been treated so far, and in vivo enrichment of MDR1-transduced cells with docetaxel treatment has been seen. There has been no apparent adverse effect from the MDR1 gene transfer.