Safety assessment of bone marrow derived MSC grown in platelet-rich plasma.

The injection of endothelial progenitor cells and mononuclear cells derived from bone marrow at the ischemic region of peripheral artery disease patients is reported to be effective for therapeutic angiogenesis; however, these cell therapies require large amounts of bone marrow to obtain sufficient numbers of cells. To solve this problem, we attempted to culture bone-marrow-derived mesenchymal stem cells (BM-MSC), which are supposed to secrete several cytokines that promote angiogenesis. We also focused on using platelet-rich plasma (PRP) as a supplement for cell culture instead of fetal bovine serum.

Enzyme-free passage of human pluripotent stem cells by controlling divalent cations.

Enzymes used for passaging human pluripotent stem cells (hPSCs) digest cell surface proteins, resulting in cell damage. Moreover, cell dissociation using divalent cation-free solutions causes apoptosis. Here we report that Mg(2+) and Ca(2+) control cell-fibronectin and cell-cell binding of hPSCs, respectively, under feeder- and serum-free culture conditions without enzyme.

Cilostazol improves lymphatic function by inducing proliferation and stabilization of lymphatic endothelial cells.

Background: Cilostazol, an inhibitor of phosphodiesterase type III, is an antiplatelet agent and vasodilator. Some clinical reports have suggested that this drug can improve progressive and refractory lymphedema.

Objective: In this study, we investigated whether cilostazol has the potential to proliferate lymphatic vessels and to improve lymphatic function using human lymphatic endothelial cells (LECs) and mouse lymphedema models.

Induction of intermediate mesoderm by retinoic acid receptor signaling from differentiating mouse embryonic stem cells.

Renal lineages including kidney are derived from intermediate mesoderm, which are differentiated from a subset of caudal undifferentiated mesoderm. The inductive mechanisms of mammalian intermediate mesoderm and renal lineages are still poorly understood. Mouse embryonic stem cells (mESCs) can be a good in vitro model to reconstitute the developmental pathway of renal lineages and to analyze the mechanisms of the sequential differentiation.

Podocalyxin is a glycoprotein ligand of the human pluripotent stem cell-specific probe rBC2LCN.

In comprehensive glycome analysis with a high-density lectin microarray, we have previously shown that the recombinant N-terminal domain of the lectin BC2L-C from Burkholderia cenocepacia (rBC2LCN) binds exclusively to undifferentiated human induced pluripotent stem (iPS) cells and embryonic stem (ES) cells but not to differentiated somatic cells. Here we demonstrate that podocalyxin, a heavily glycosylated type 1 transmembrane protein, is a glycoprotein ligand of rBC2LCN on human iPS cells and ES cells. When analyzed by DNA microarray, podocalyxin was found to be highly expressed in both iPS cells and ES cells.

Reduction of N-glycolylneuraminic acid in human induced pluripotent stem cells generated or cultured under feeder- and serum-free defined conditions.

Background: The successful establishment of human induced pluripotent stem cells (hiPSCs) has increased the possible applications of stem cell research in biology and medicine. In particular, hiPSCs are a promising source of cells for regenerative medicine and pharmacology. However, one of the major obstacles to such uses for hiPSCs is the risk of contamination from undefined pathogens in conventional culture conditions that use serum replacement and mouse embryonic fibroblasts as feeder cells.

In vitro organogenesis from undifferentiated cells in Xenopus.

Amphibians have been used for over a century as experimental animals. In the field of developmental biology in particular, much knowledge has been accumulated from studies on amphibians, mainly because they are easy to observe and handle. Xenopus laevis is one of the most intensely investigated amphibians in developmental biology at the molecular level.

Isolation and differentiation of Xenopus animal cap cells.

Xenopus is used as a model animal for investigating the inductive events and organogenesis that occur during early vertebrate development. Given that they are easy to obtain in high numbers and are relatively large in size, Xenopus embryos are excellent specimens for performing manipulations such as microinjection and microsurgery. The animal cap, which is the area around the animal pole of the blastula, is destined to form the ectoderm during normal development.

Ripply2 is essential for precise somite formation during mouse early development.

The regions of expression of Ripply1 and Ripply2, presumptive transcriptional corepressors, overlap at the presomitic mesoderm during somitogenesis in mouse and zebrafish. Ripply1 is required for somite segmentation in zebrafish, but the developmental role of Ripply2 remains unclear in both species. Here, we generated Ripply2 knock-out mice to investigate the role of Ripply2.

Dullard promotes degradation and dephosphorylation of BMP receptors and is required for neural induction.

Bone morphogenetic proteins (BMPs) regulate multiple biological processes, including cellular proliferation, adhesion, differentiation, and early development. In Xenopus development, inhibition of the BMP pathway is essential for neural induction. Here, we report that dullard, a gene involved in neural development, functions as a negative regulator of BMP signaling.

Ledgerline, a novel Xenopus laevis gene, regulates differentiation of presomitic mesoderm during somitogenesis.

Segmentation of the vertebrate body via the sequential formation of somites is an important process in embryogenesis. This sequential process is governed by the activation and regulation of Notch-related molecular oscillators by fibroblast growth factor and retinoic acid (RA) signaling. In this study, we identified ledgerline, a novel gene of Xenopus laevis expressed specifically in the presomitic mesoderm.

Growing kidney in the frog.

An understanding of the regulation of kidney development has increased dramatically in the past decade. The pronephros, mesonephros, and metanephros represent three distinct renal organs that function, in succession, as the vertebrate excretory system during development of the kidney. These three organ systems are derived from the intermediate mesoderm and develop in a well-defined temporal and spatial sequence.

The role of XTRAP-gamma in Xenopus pronephros development.

We isolated and characterized the Xenopus translocon-associated protein XTRAP-gamma, one of four subunits of the translocon-associated protein complex. TRAP has been proposed to aid the translocation of nascent polypeptides into the lumen of the endoplasmic reticulum, but this has not been demonstrated until now. XTRAP-gamma was specifically expressed in the pronephros tubules of Xenopus laevis from stage 25 during kidney development.

The role of Xenopus frizzled-8 in pronephric development.

Vertebrates use two or three forms of kidney successively during development and the nephric duct is essential for this succession of kidney induction. While transcripts of many Wnt ligands and Wnt receptor Frizzled genes have been localized in developing kidney, the relationship between Wnt signaling and nephric duct development remains unknown. This study investigated the role of Xenopus frizzled-8 (Xfz8) in pronephros development.

The isolation and characterization of XC3H-3b: a CCCH zinc-finger protein required for pronephros development.

We describe the isolation and characterization of the RNA-binding protein XC3H-3b that is expressed during pronephros development. XC3H-3b is a member of the TTP/TIS family of CCCH tandem zinc-finger proteins, which are physiological stimulators of instability for the mRNA encoding tumor necrosis factor-alpha in certain cell types. XC3H-3b is localized primarily to the mesodermal tissues around the pronephros.

Molecular cloning and characterization of dullard: a novel gene required for neural development.

In a screen for genes expressed in neural tissues and pronephroi, we isolated a novel gene, named dullard. Dullard protein contains the C-terminal conserved domain of NLI-IF (Nuclear LIM Interactor-Interacting Factor), a protein whose function is not yet characterized. Dullard mRNA was maternally derived and localized to the animal hemisphere.