Dermal papilla cells serially cultured with Wnt-10b sustain their hair follicle induction activity after transplantation into nude mice.

Dermal papilla (DP) cells are associated with the development of hair follicles (HFs) and regulation of the hair cycle. However, primary DP cells prepared from cultured HFs are known to lose their ability to induce HF after culturing in standard media, for example, fibroblast growth conditions. We explored a new culture condition by which DP cells maintained their HF induction ability.

Environmental stimulation changes tissue-type plasminogen activator activity in the adult mouse hippocampus.

Tissue-type plasminogen activator (tPA) plays an important role in synaptic plasticity and contributes to several brain functions such as memory, learning, and behaviours. Although a number of studies have demonstrated that various kinds of stimuli such as electrophysiological stimulation, excitotoxic injury, and stress change tPA activity in the brain, no studies have ever examined whether environmental stimulation affect tPA activity in the brain. The aim of this study is to clarify the effect of environmental enrichment on tPA activity in the hippocampus and cerebral cortex of adult mouse brain.

Modulated differentiation of embryonic stem cells into hepatocyte-like cells by coculture with hepatic stellate cells.

We investigated the effects of coculture with hepatic stellate cells (HSCs) on the differentiation of embryonic stem (ES) cells and embryoid bodies (EBs). Rat HSCs were incubated until becoming semi-confluent and adherent to the dish. Undifferentiated mouse ES cells and 4-day EBs were cultured in gelatin-coated or HSC-feeder dishes, then induced hepatocyte-like cells and the remaining undifferentiated ES cells were examined using immunocytochemical and RT-PCR methods.

Isolation and characterization of murine hepatocytes following collagenase infusion into left ventricle of heart.

A method for obtaining mouse hepatocytes by infusing collagenase solution into the left ventricle was established. This technique was shown to be equivalent to the intra-portal infusion method and more practical, especially in postnatal mice with a small body size.

Effects of Wnt-10b on proliferation and differentiation of adult murine skin-derived CD34 and CD49f double-positive cells.

Although mouse Wnt-10b has been shown to play various roles in a wide range of biological actions, the effects on epithelial stem/progenitor cells in the skin have not been reported. In the present study, we investigated the effects of Wnt-10b on proliferation and differentiation of murine skin-derived CD34 and CD49f double-positive (CD34(+)CD49f(+)) cells, a supposed fraction as enriched epithelial stem/progenitor cells. The cells were prepared from dorsal skin samples obtained from young adult mice as alpha6 integrin (CD49f) and CD34 double-positive cells by fluorescent activated cell sorting (FACS), and they were cultured with or without Wnt-10b to investigate its effects on proliferation and differentiation.

Cotransplantation of mouse embryonic stem cells and bone marrow stromal cells following spinal cord injury suppresses tumor development.

Embryonic stem (ES) cells are a potential source for treatment of spinal cord injury (SCI). Although one of the main problems of ES cell-based cell therapy is tumor formation, there is no ideal method to suppress tumor development. In this study, we examined whether transplantation with bone marrow stromal cells (BMSCs) prevented tumor formation in SCI model mice that received ES cell-derived grafts containing both undifferentiated ES cells and neural stem cells.

Treatment of Parkinson's disease model mice with allogeneic embryonic stem cells: necessity of immunosuppressive treatment for sustained improvement.

Objective: The purpose of the present study was to examine the efficacy of transplantation of mouse embryonic stem (ES) into Parkinson's disease (PD) model mice as well as the necessity of immunosuppression in allogeneic donor-host combinations.

Materials And Methods: ES cells, derived from SvJ129 strain mice, were differentiated into tyrosine hydroxylase (TH)-positive neurons in vitro by an embryoid body (EB)-based multistep differentiation method and used as graft cells for PD mice, which were prepared by injection of 6-hydroxydopamine (OHDA) into C57BL/6, BALB/c and C3H/HeN strains. Mice from each strain were divided into Groups 1-3.

Embryonic stem cells reduce liver fibrosis in CCl4-treated mice.

We transplanted undifferentiated embryonic stem (ES) cells into the spleens of carbon tetrachloride (CCl(4))-treated mice to determine their effects on liver fibrosis. Carbon tetrachloride at 0.5 ml/kg of body weight was injected intraperitoneally into C57BL/6 mice twice weekly for up to 20 weeks.

Cotransplantation of Mouse Embryonic Stem Cells and Bone Marrow Stromal Cells following Spinal Cord Injury Suppresses Tumor Development.

Embryonic stem (ES) cells are a potential source for treatment of spinal cord injury (SCI). Although one of the main problems of ES cell-based cell therapy is tumor formation, there is no ideal method to suppress tumor development. In this study, we examined whether transplantation with bone marrow stromal cells (BMSCs) prevented tumor formation in SCI model mice that received ES cell-derived grafts containing both undifferentiated ES cells and neural stem cells.

Localization of chondroitin sulfate proteoglycan versican in adult brain with special reference to large projection neurons.

Versican is a chondroitin sulfate proteoglycan belonging to the lectican family. Versican has two glycosaminoglycan attachment regions, named the GAG alpha and GAG beta domains, which are both regulated by alternative splicing and yield four protein isoforms. We have investigated the expression and localization of versican in the developing and adult brain by using anti-versican GAG alpha and GAG beta antibodies.

CoCl(2) inhibits neural differentiation of retinoic acid-treated embryoid bodies.

The effects of CoCl(2) on retinoic acid (RA)-treated embryoid bodies (EBs) were investigated. Four-day EBs were treated with 5x10(-6) M of RA for 4 d, then subjected to attached culturing for 7 d in the presence of CoCl(2) at 0, 20, and 100 microM. Differentiation into MAP2- and GFAP-immunopositive cells was inhibited by CoCl(2) in a dose-dependent manner.

A familial case of visceral toxocariasis due to consumption of raw bovine liver.

We present 3 adult cases of visceral toxocariasis from the same family, who each consumed thin slices of raw bovine liver weekly, and developed eosinophilia and multiple small lesions in their livers and lungs. Serological examinations using the larval excretory-secretory product of Toxocara canis strongly indicated infection with Toxocara species larvae. The patients responded well to treatment with albendazole.

Diagnostic problems in a patient with amicrofilaremic Loa loa.

We report a Japanese patient with loiasis who became infected in Cameroon. Despite the clinical history and laboratory data providing adequate evidence for suspecting loiasis, microfilariae were not detected in the blood. It is important to note that most infected travelers whose home countries are in nonendemic regions are amicrofilaremic.

Wnt-10b, uniquely among Wnts, promotes epithelial differentiation and shaft growth.

Although Wnts are expressed in hair follicles throughout life from embryo to adult, and considered to be critical for their development and maturation, their roles remain largely unknown. In the present study, we investigated the effects of Wnts (Wnt-3a, Wnt-5a, Wnt-10b, and Wnt-11) on epithelial cell differentiation using adult mouse-derived primary skin epithelial cell (MPSEC) cultures and hair growth using hair follicle organ cultures. Only Wnt-10b showed evident promotion of epithelial cell differentiation and hair shaft growth, in contrast to Wnt-3a, 5a, and 11.

DACS, novel matrix structure composed of chondroitin sulfate proteoglycan in the brain.

Chondroitin sulfate proteoglycans (CSPGs) are major components of the extracellular matrix (ECM) in the brain. In the adult cerebral cortex, there are special CSPG-containing structures known as perineuronal nets (PNNs), which are highly condensed ECM structures. Here, we report a novel CSPG-containing structure distinct from PNNs in the adult mouse cerebral cortex.

Effects of Wnt-10b on hair shaft growth in hair follicle cultures.

Wnts are deeply involved in the proliferation and differentiation of skin epithelial cells. We previously reported the differentiation of cultured primary skin epithelial cells toward hair shaft and inner root sheath (IRS) of the hair follicle via beta-catenin stabilization caused by Wnt-10b, however, the effects of Wnt-10b on cultured hair follicles have not been reported. In the present study, we examined the effects of Wnt-10b on shaft growth using organ cultures of whisker hair follicles in serum-free conditions.

Cystic echinococcosis in immigrant from Peru: first case treated with percutaneous treatment in Japan.

We report a case of cystic echinococcosis (CE) caused by Echinococcusgranulosus, for which a modified percutaneous evacuation (PEVAC) treatment was applied. The patient had immigrated from Peru to Japan and had 2 hydatid cystic masses, 1 located in segment (S)5 of the liver and the other in S3 (5.3 and 3.

Embryonic stem cells develop into hepatocytes after intrasplenic transplantation in CCl4-treated mice.

Aim: To transplant undifferentiated embryonic stem (ES) cells into the spleens of carbon tetrachloride (CCl4)-treated mice to determine their ability to differentiate into hepatocytes in the liver.

Methods: CCl4, 0.5 mL/kg body weight, was injected into the peritoneum of C57BL/6 mice twice a week for 5 wk.

Promoted differentiation of cynomolgus monkey ES cells into hepatocyte-like cells by co-culture with mouse fetal liver-derived cells.

Aim: To explore whether a co-culture of cynomolgus monkey embryonic stem (cES) cells with embryonic liver cells could promote their differentiation into hepatocytes.

Methods: Mouse fetal liver-derived cells (MFLCs) were prepared as adherent cells from mouse embryos on embryonic d (ED) 14, after which undifferentiated cES cells were co-cultured with MFLCs. The induction of cES cells along a hepatic lineage was examined in MFLC-assisted differentiation, spontaneous differentiation, and growth factors (GF) and chemicals-induced differentiations (GF-induced differentiation) using retinoic acid, leukemia inhibitory factor (LIF), FGF2, FGF4, hepatocyte growth factor (HGF), oncostatin M (OSM), and dexamethasone.

Cynomolgus monkey embryonic stem cell lines express green fluorescent protein.

We successfully established cynomolgus monkey embryonic stem (cES) cell lines expressing enhanced green fluorescent protein (GFP) by introducing a GFP-encoding gene under cytomegalovirus immediate early enhancer (CMVIE) promoter regulation into cES cells. The cells maintained the ability of in vitro differentiation toward ectodermal, mesodermal, and endodermal lineages, and produced teratomas composed of tissues derived from the three embryonic germ layers when transplanted into severe combined immunodeficient disease mice. GFP expression was also observed in the differentiated cells.

Derivation and characterization of hepatocytes from embryonic stem cells in vitro.

From a therapeutic perspective on liver, the use of embryonic stem (ES) technology in the generation of a large number of high-functional hepatocytes developed from ES cells for cell transplantation is anticipated. We have explored a three-dimensional culture system in which hepatocytes differentiated from mouse ES cells by transfection with the hepatocyte nuclear factor-3beta possess high metabolic functions that can be maintained long term.

Use of differentiating embryonic stem cells in the Parkinsonian mouse model.

Progressive loss of dopaminergic neurons in the substantia nigra pars compacta and the following reduction in striatal dopamine cause Parkinson's disease (PD). Transplantation of dopamine-producing cells into the striatum is a proposed treatment modality. In this report, we describe a model experiment assessing the effectiveness of mouse embryonic stem (ES) cell-derived dopaminergic neurons using a mouse model of PD.