The potential of stem cells in adult tissues representative of the three germ layers.

Mature adult tissues contain stem cells that express many genes normally associated with the early stage of embryonic development, when maintained in appropriate environments. Cells procured from adult tissues representative of the three germ layers (spinal cord, muscle, and lung), each exhibiting the potential to mature into cells representative of all three germ layers. Cells isolated from adult tissues of different germ layer origin were propagated as nonadherent clusters or spheres that were composed of heterogeneous populations of cells.

Differences in stress response after UVC or UVA irradiation in Vibrio parahaemolyticus.

The SOS response is a global regulatory network for repairing DNA damage induced by various environmental stresses such as UV irradiation. The Escherichia coli SOS response has been extensively studied. However, there are no reports on the SOS response in Vibrio parahaemolyticus.

In vivo evaluation of novel nitroxyl radicals with reduction stability.

Nitroxyl radicals (nitroxide) have great potential advantages as spin probes, antioxidants, contrast agents, and radiation-protecting agents. However, they are readily reduced by reductants in cells and lose their paramagnetic nature. Recently, tetraethyl-substituted nitroxyl radicals have been reported to have high stability toward reduction by ascorbic acid (AsA).

Portable microcontact printing device for cell culture.

In this work, we developed a portable device to perform microcontact printing in a safety cabinet for cell culture. The device was designed to be small and non-bulky, easy to sterilize, while not requiring the use of electricity, and which requires very little manual handling. Moreover, the portable microcontact printer is reproducibly fabricated with a rapid prototyping system, and allows for the easy micropatterning of biomolecules with a resolution ranging from 20 to 500 μm.

Preparation of keratinocyte culture medium for the clinical applications of regenerative medicine.

Keratinocyte culture medium (KCM) has been used for the in vitro culture of keratinocytes and other types of epithelial cells, and the medium includes various ingredients. In this study, two modified KCMs were prepared. In the first, insulin, hydrocortisone and antibiotics that are normally included in KCM were replaced with clinically approved pharmaceutical agents, except transferrin and selenium; in the second, cholera toxin (CT) was replaced by L-isoproterenol (ISO).

In vivo 3D analysis with micro-computed tomography of rat calvaria bone regeneration using periosteal cell sheets fabricated on temperature-responsive culture dishes.

Transplantable cell sheets containing osteoblasts were fabricated from periostea on temperature-responsive culture dishes. This study demonstrated the time-course of bone regeneration in living small animals. This continuous observation of bone regeneration was achieved by micro-computed tomography (µCT), which assessed the osteogenic capability of periosteal cells without biodegradable scaffolds.

Surgical anatomy of the swine face.

In order to develop tissue engineering applications for oral and maxillofacial surgery, the surgical anatomy of the miniature pig's face was investigated in three miniature pig cadavers and three anaesthetized miniature pigs using identical procedures that were previously described for humans. A preauricular incision with a retromandibular and a submandibular extension was initially made through the facial skin and subcutaneous tissues. The underlying tissues were then carefully dissected in order to progressively expose the platysma muscle, the superficial layer of deep cervical fascia, the marginal mandibular branch and buccal branch of the facial nerve, the mental nerve and the mandibular skeleton.

Three-dimensional cell-dense constructs containing endothelial cell-networks are an effective tool for in vivo and in vitro vascular biology research.

Angiogenesis is a complicated natural process, and understanding the mechanism by which it occurs is important for medical, pharmaceutical, and cell biological sciences. Many techniques for investigating angiogenesis have been reported. In this study, we introduced a novel application of a cell culture technique that can be used in in vitro and in vivo vascular biology research.

Comb-type grafted poly(N-isopropylacrylamide) gel modified surfaces for rapid detachment of cell sheet.

A comb-type grafted poly(N-isopropylacrylamide) (PIPAAm) gel modified surface was newly developed for providing a rapid cell sheet recovery for tissue engineering. PIPAAm macromonomer was prepared by the etherification reaction of the hydroxyl terminal moieties of PIPAAm with acryloyl chloride, followed by the radical telomerization reaction of N-isopropylacrylamide (IPAAm) monomer using 2-mercaptoethanol as a chain transfer agent. Solution containing IPAAm monomer and PIPAAm macromonomer was spread on the surface of tissue culture polystyrene (TCPS), and then the surface was subjected to electron beam irradiation for grafting the monomer and macromonomer on the surfaces, resulting in comb-type grafted PIPAAm gel modified TCPS (GG-TCPS).

Assessment of cell sheets derived from human periodontal ligament cells: a pre-clinical study.

Periodontal-ligament-derived cells (PDL cells) have stem-cell-like properties and, when implanted into periodontal defects in vivo, can induce periodontal regeneration including the formation of new bone, cementum, and periodontal ligament. We have previously demonstrated that PDL cell sheets, harvested from temperature-responsive cell culture dishes, have a great potential for periodontal regeneration. The purpose of this study has been to validate the safety and efficacy of human PDL (hPDL) cell sheets for use in clinical trials.

Validation of human periodontal ligament-derived cells as a reliable source for cytotherapeutic use.

Aim: Periodontal ligament (PDL) is a reliable cell source for periodontal regeneration. In this study, an optimal protocol for the extraction, expansion, and characterization of human PDL (hPDL) cells was examined for clinical trials.

Materials And Methods: hPDL tissues were obtained from 41 surgically extracted teeth and digested with enzymes.

Anisotropic mechanical properties of collagen hydrogels induced by uniaxial-flow for ocular applications.

Engineering of well-organized tissue constructs is active in the field of material science for biomedical applications. Here, we propose a method for orienting collagen in transparent high-density collagen hydrogels using a simple rolling method. Structural organization and mechanical function were adjusted by regulating the thickness of the construct and the cross-linking reagent.

Controlled chain length and graft density of thermoresponsive polymer brushes for optimizing cell sheet harvest.

Novel thermoresponsive polymer brush surfaces for harvesting cell sheet were fabricated by the surface-initiated RAFT polymerization of N-isopropylacrylamide (IPAAm) on azoinitiator-immobilized glass substrates in the presence of dithiobenzoate compound as a chain transfer agent (CTA). The chain length of the grafted PIPAAm on the surface was controlled by changing CTA concentration. Additionally, PIPAAm graft density on the surface was successfully regulated by grafting from azoinitiator-immobilized surfaces with various densities.

A novel method of culturing human oral mucosal epithelial cell sheet using post-mitotic human dermal fibroblast feeder cells and modified keratinocyte culture medium for ocular surface reconstruction.

Background/aims: To cultivate human oral mucosal epithelial cell sheets with post-mitotic human dermal fibroblast feeder cells and modified keratinocyte culture medium for ocular surface reconstruction.

Methods: Human oral mucosal epithelial cells obtained from three healthy volunteers were cultured with x-ray-treated dermal fibroblasts (fibroblast group) and NIH/3T3 feeder layers (3T3 group) on temperature-responsive culture dishes. Media were supplemented using clinically approved products.

Characterization of ultra-thin temperature-responsive polymer layer and its polymer thickness dependency on cell attachment/detachment properties.

Ultra thin poly(N-isopropylacrylamide) (PIPAAm) modified glass coverslips (PIAPAm-CS) using electron beam irradiation exhibited a clear relationship between the polymer thickness and thermal cell adhesion/detachment behavior. The polymer thickness dependency and the characteristic of ultra thin PIPAAm layer, has been illustrated in terms of the molecular motion of the modified PIPAAm chains. PIPAAm-CSs surfaces with various area-polymer densities and thicknesses were characterized by AFM and protein adsorption assay.

A protocol for immunofluorescence staining of floating neurospheres.

This protocol describes the immunofluorescence staining of floating neurospheres in culture plates. Although this protocol is similar to conventional immunofluorescence staining, the staining procedure of floating neurospheres in multiwell culture plates and the washing procedure are different. Neurospheres in culture plates are transferred to a 12-well plate using a 200-1000microL pipette.

Cell attachment-detachment control on temperature-responsive thin surfaces for novel tissue engineering.

Temperature-responsive intelligent surfaces, prepared by the modification of an interface mainly with poly(N-isopropylacrylamide) and its derivatives, have been investigated. Such surfaces exhibit temperature-responsive hydrophilic/hydrophobic alterations with external temperature changes, which, in turn, result in thermally modulated attachment and detachment with cells. The advantage of this system is that cells cultured on such temperature-responsive surfaces can be recovered as single cells and/or confluent cell sheets, while keeping the deposited extracellular matrix intact, simply by lowering the temperature without conventional enzymatic treatment.

A new colored beverage disinfection system using UV-A light-emitting diodes.

In this study we evaluated the ability of the UV-A-LED to eliminate bacteria in a colored beverage. Ten edible pigments were used to make a colored solution at concentrations of 1.0%, 0.

Pre-vascularization of in vitro three-dimensional tissues created by cell sheet engineering.

Reconstructing a vascular network is a common task for three-dimensional (3-D) tissue engineering. Three-dimensional stratified tissues were created by stacking cell sheets, and the co-culture with endothelial cells (ECs) in the tissues was found to lead to in vitro pre-vascular network formation and promoted in vivo neovascularization after their transplantation. In this study, to clarify the effect of tissue fabrication process on a pre-vascular network formation, human origin ECs were introduced into the stratified tissue in several different ways, and the behavior of ECs in various positions of the 3-D tissue were compared each other.

Taxonomic reconsideration of a sequestrate fungus, Octaviania columellifera, with the proposal of a new genus, Heliogaster, and its phylogenetic relationships in the Boletales.

During taxonomic revision of genus Octaviania in Japan we examined herbarium and fresh specimens of O. columellifera and O. asterosperma sensu S.

E-cadherin and claudin-4 expression has circadian rhythm in adult rat kidney.

Background: Urinary sodium excretion varies during the day. It is unknown whether expression levels of cell-cell junctions in the kidney are dynamic and associated with urinary sodium excretion.

Methods: Adult Sprague Dawley rats were fed ad libitum or exclusively during the day, or kept under fasting condition.

Cell sheet engineering: a unique nanotechnology for scaffold-free tissue reconstruction with clinical applications in regenerative medicine.

Cell sheet technology (CST) is based on the use of thermoresponsive polymers, poly(N-isopropylacrylamide) (PIPAAm). The surface of PIPAAms is formulated in such a way as to make its typical thickness <100 nm. In this review, we first focus on how the methods of PIPAAm-grafted surface preparations and functionalization are important to be able to harvest a functional cell sheet, to be further transplanted.

Silkworm pupae (Bombyx mori) are new sources of high quality protein and lipid.

This study was performed to evaluate the nutritional value of silkworm pupae (Bombyx mori) and the content of α-glucosidase inhibitor. The percentages of total protein and lipid contents by dry weight were 55.6 and 32.

Determination of reactive oxygen species associated with the degeneration of dopaminergic neurons during dopamine metabolism.

Oxidative stress is believed to be an important mechanism underlying dopamine-induced neuronal damage. This study provides X-band electron spin resonance (ESR) spectroscopic evidence for reactive oxygen species (ROS) generation during dopamine metabolism. The authors induced excess dopamine metabolism in the mouse striatum by bathing it in tyramine-containing perfusate using microdialysis.