Somatic stem cells have been isolated from multiple human tissues for their potential usefulness in cell therapy. Currently, mesenchymal stromal cells (MSCs) are prepared after several passages requiring a few months of cell culture. In this study, we used a prospective isolation method of somatic stem cells from gestational or fat tissues, which were identified using CD73 antibody. CD73-positive population from various tissues existed individually in flowcytometric pattern, especially subcutaneous fat- and amniotic-derived cells showed the highest enrichment of CD73-positive cells. Moreover, the cell populations isolated with the prospective method showed higher proliferative capacity and stem cell marker expression, compared to the cell populations which isolated through several passages of culturing whole living cells: which we named “conventional method” in this paper. Furthermore, the therapeutic potential of CD73-positive cells was evaluated in vivo using a mouse model of pulmonary fibrosis. After intranasal administration, murine CD73-positive cells reduced macrophage infiltration and inhibited fibrosis development. These results suggest that further testing using CD73-positive cells may be beneficial to help establish the place in regenerative medicine use.

As a constituent of polysaccharides and glycoconjugates, β-d-galactofuranose (Galf) exists in several pathogenic microorganisms. Although we recently identified a β-d-galactofuranosidase (Galf-ase) gene, ORF1110, in the Streptomyces strain JHA19, very little is known about the Galf-ase gene. Here, we characterized a strain, named JHA26, in the culture supernatant of which exhibited Galf-ase activity for 4-nitrophenyl β-d-galactofuranoside (pNP-β-d-Galf) as a substrate. Draft genome sequencing of the JHA26 strain revealed a putative gene, termed ORF0643, that encodes Galf-ase containing a PA14 domain, which is thought to function in substrate recognition. The recombinant protein expressed in Escherichia coli showed the Galf-specific Galf-ase activity and also released galactose residue of the polysaccharide galactomannan prepared from Aspergillus fumigatus, suggesting that this enzyme is an exo-type Galf-ase. BLAST searches using the amino acid sequences of ORF0643 and ORF1110 Galf-ases revealed two types of Galf-ases in Actinobacteria, suggesting that Galf-specific Galf-ases may exhibit discrete substrate specificities.

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1. 734-85531-2-3There are certain transporters that function effectively even at ice-cold temperatures, such as facilitative glucose transporter (GLUT) 1 and equilibrative nucleoside transporter (ENT) 1 in human erythrocyte membranes. In order to evaluate the characteristics of these transporters precisely, inhibition of substrate efflux during stopping and washing processes is essential. In this study, we attempted to develop a novel stop solution that can be used universally for membrane transport studies. As candidate compounds for preparing a novel stop solution, protein denaturants such as urea, 2-mercaptoethanol, formaldehyde, and glutaraldehyde were used. D-Glucose and uridine were used as substrates for GLUT1 and ENT1, respectively. [ 3 H]D-Glucose taken up by rightside-out erythrocyte membrane vesicles (ROVs) was almost completely effluxed during washing with ice-cold stop solution without phloretin.Protein denaturants inhibited the efflux of these substrates, and increased the remaining amount of substrates in ROVs. Especially, the combination of urea/formaldehyde or 2-mercaptoethanol/glutaraldehyde was effective as a stop solution for both GLUT1-and ENT1-mediated efflux. In addition, these stop solutions were applicable for the transport study in cultured HepG2 cells. Our results indicate that these novel stop solutions can be used universally for membrane transport studies with isolated membrane vesicles and culture cells.Corresponding Author