Neural stem cell (NSC) transplantation has been proposed as a future therapy for neurodegenerative disorders. However, NSC transplantation will be hampered by the limited number of brain donors and the toxicity of immunosuppressive regimens that might be needed with allogeneic transplantation. These limitations may be avoided if NSCs can be generated from clinically accessible sources, such as bone marrow (BM) and peripheral blood samples, that are suitable for autologous transplantation. We report here that NSCs can be generated from human BM-derived mesenchymal stem cells (MSCs). When cultured in NSC culture conditions, 8% of MSCs were able to generate neurospheres. These MSC-derived neurospheres expressed characteristic NSC antigens, such as nestin and musashi-1, and were capable of self-renewal and multilineage differentiation into neurons, astrocytes, and oligodendrocytes. Furthermore, when these MSC-derived neurospheres were cocultured with primary astrocytes, they differentiate into neurons that possess both dendritic and axonal processes, form synapses, and are able to fi re tetrodotoxin-sensitive action potentials. When these MSC-derived NSCs were switched back to MSC culture conditions, a small fraction of NSCs (averaging 4-5%) adhered to the culture fl asks, proliferated, and displayed the morphology of MSCs. Those adherent cells expressed the characteristic MSC antigens and regained the ability to differentiate into multiple mesodermal lineages. Data presented in this study suggest that MSCs contain a small fraction (averaging 4-5%) of a bipotential stem cell population that is able to generate either MSCs or NSCs depending on the culture conditions.

The hematopoietic stem cell (HSC) compartment is composed of long-term reconstituting (LTR) and short-term reconstituting (STR) stem cells. LTR HSC can reconstitute the hematopoietic system for life, whereas STR HSC can sustain hematopoiesis for only a few weeks in the mouse. Several excellent gene expression profiles have been obtained of the total hematopoietic stem cell population. We have used five-color FACS sorting to isolate separate populations of LTR and STR stem cell subsets. The LTR HSC has the phenotype defined as Lin ؊ Sca ؉ Kit ؉ 38 ؉ 34 ؊ ; two subsets of STR HSC were obtained with phenotypes of Lin ؊ Sca ؉ Kit ؉ 38 ؉ 34 ؉ and Lin ؊ Sca ؉ Kit ؉ 38 ؊ 34 ؉ . The microarray profiling study reported here was able to identify genes specific for LTR functions. In the interrogated genes (Ϸ12,000 probe sets corresponding to 8,000 genes), 210 genes are differentially expressed, and 72 genes are associated with LTR activity, including membrane proteins, signal transduction molecules, and transcription factors. Hierarchical clustering of the 210 differentially expressed genes suggested that they are not bone marrow-specific but rather appear to be stem cell-specific. Transcription factor-binding site analysis suggested that GATA3 might play an important role in the biology of LTR HSC.microarray ͉ regulation

A chimeric retroviral vector (33E67) containing a CD33-specific single-chain antibody was generated in an attempt to target cells displaying the CD33 surface antigen. The chimeric envelope protein was translated, processed, and incorporated into viral particles as efficiently as wild-type envelope protein. The viral particles carrying the 33E67 envelope protein could bind efficiently to the CD33 receptor on target cells and were internalized, but no gene transfer occurred. A unique experimental approach was used to examine the basis for this postbinding block. Our data indicate that the chimeric envelope protein itself cannot participate in the fusion process, the most reasonable explanation being that this chimeric protein cannot undergo the appropriate conformational change that is thought to be triggered by receptor binding, a suggested prerequisite to subsequent fusion and core entry. These results indicate that the block to gene transfer in this system, and probably in most of the current chimeric retroviral vectors to date, is the inability of the chimeric envelope protein to undergo this obligatory conformational change.

STEM CELLS 2006;24: 1549 -1555

The envelope glycoproteins of the mammalian type C retroviruses consist of two subunits, a surface (SU) protein and a transmembrane (TM) protein. SU binds to the viral receptor and is thought to trigger conformational changes in the associated TM protein that ultimately lead to the fusion of viral and host cell membranes. For Moloney murine leukemia virus (MoMuLV), the envelope protein probably exists as a trimer. We have previously demonstrated that the coexpression of envelope proteins that are individually defective in either the SU or TM subunits can lead to functional complementation (Y. Zhao et al., J. Virol. 71:6967–6972, 1997). We have now extended these studies to investigate the abilities of a panel of fusion-defective TM mutants to complement each other. This analysis identified distinct complementation groups within TM, with implications for interactions between different regions of TM in the fusion process. In viral particles, the C-terminal 16 amino acids of the MoMuLV TM (the R peptide) are cleaved by the viral protease, resulting in an increased fusogenicity of the envelope protein. We have examined the consequences of R peptide cleavage for the different TM fusion mutants and have found that this enhancement of fusogenicity can only occur incis to certain of the TM mutants. These results suggest that R peptide cleavage enhances the fusogenicity of the envelope protein by influencing the interaction of two distinct regions in the TM ectodomain.

These data support that GLP-1 directs human ES cell differentiation into insulin-producing cells via hedgehog, cyclic adenosine monophosphate, and phosphatidylinositol-3-kinase pathways.

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IntroductionCardiac hypertrophy is an independent risk factor for heart failure. However, the underlying mechanisms of cardiac hypertrophy are still unclear. Nintedanib is a Food and Drug Administration (FDA) approved therapeutic agent for the treatment of progressive fibrosing lung diseases.Material and methodsIn this study, we examined the effects of nintedanib on cardiac hypertrophy using an in vivo murine model with the transverse aortic constriction (TAC) operation and an in vitro cardiomyocytes model stimulated with Ang II.ResultsNintedanib has a protective effect on cardiac function in TAC mice with decreased heart rates, heart weight/body weight (HW/BW), and reduced plasma levels of creatine kinase-MB (CK-MB) and aspartate aminotransferase (AST). Wheat germ agglutinin (WGA) staining proved that the increased cardiomyocytes sizes in TAC mice were restored by nintedanib treatment. Nintedanib also reversed the decreased plasma levels of oxidative markers nuclear factor erythroid-2-related factor 2 (Nrf2), lipid peroxidation products thiobarbituric acid reactive substances (TBARS), and GSH, as well the increased homocysteine (Hcy) levels in TAC mice. In the in vitro cardiomyocytes model, cells were treated with nintedanib, followed by Ang II stimulation. Nintedanib improved Ang II induction-caused cell injury and oxidative stress in H9C2 cells, as shown by the decreased release of lactate dehydrogenase (LDH), and elevated mRNA levels of GPX1 and HO-1. Mechanistically, we prove that the protective effect of nintedanib is mediated by SIRT1.ConclusionsIn conclusion, this study demonstrates the protective effects of nintedanib on cardiac hypertrophy both in vivo and in vitro, which was attributed to its anti-oxidative activity through regulating SIRT1 expression.