23 research outputs found
Purification of functional human ES and iPSC-derived midbrain dopaminergic progenitors using LRTM1
Human induced pluripotent stem cells (iPSCs) can provide a promising source of midbrain dopaminergic (mDA) neurons for cell replacement therapy for Parkinson's disease (PD). However, iPSC-derived donor cells inevitably contain tumorigenic or inappropriate cells. To eliminate these unwanted cells, cell sorting using antibodies for specific markers such as CORIN or ALCAM has been developed, but neither marker is specific for ventral midbrain. Here we employ a double selection strategy for cells expressing both CORIN and LMX1A::GFP, and report a cell surface marker to enrich mDA progenitors, LRTM1. When transplanted into 6-OHDA-lesioned rats, human iPSC-derived LRTM1 + cells survive and differentiate into mDA neurons in vivo, resulting in a significant improvement in motor behaviour without tumour formation. In addition, there was marked survival of mDA neurons following transplantation of LRTM1 + cells into the brain of an MPTP-treated monkey. Thus, LRTM1 may provide a tool for efficient and safe cell therapy for PD patients
Physiological and Genomic Features of a Novel Sulfur-Oxidizing Gammaproteobacterium Belonging to a Previously Uncultivated Symbiotic Lineage Isolated from a Hydrothermal Vent
<div><p>Strain Hiromi 1, a sulfur-oxidizing gammaproteobacterium was isolated from a hydrothermal vent chimney in the Okinawa Trough and represents a novel genus that may include a phylogenetic group found as endosymbionts of deep-sea gastropods. The SSU rRNA gene sequence similarity between strain Hiromi 1 and the gastropod endosymbionts was approximately 97%. The strain was shown to grow both chemolithoautotrophically and chemolithoheterotrophically with an energy metabolism of sulfur oxidation and O<sub>2</sub> or nitrate reduction. Under chemolithoheterotrophic growth conditions, the strain utilized organic acids and proteinaceous compounds as the carbon and/or nitrogen sources but not the energy source. Various sugars did not support growth as a sole carbon source. The observation of chemolithoheterotrophy in this strain is in line with metagenomic analyses of endosymbionts suggesting the occurrence of chemolithoheterotrophy in gammaproteobacterial symbionts. Chemolithoheterotrophy and the presence of homologous genes for virulence- and quorum sensing-related functions suggest that the sulfur-oxidizing chomolithotrophic microbes seek animal bodies and microbial biofilm formation to obtain supplemental organic carbons in hydrothermal ecosystems.</p></div
Predicted sulfur oxidation pathways of strain Hiromi 1 inferred from the previously published literatures [63], [66]–[68], [74], [75].
<p>TCEP; tris(2-carboxyethyl)phosphine.</p
Predicted central metabolism of strain Hiromi 1.
<p>Red lines indicate conserved pathways in most of the publically accessible genomes of the chemolithoautotrophic <i>Gammaproteobacteria</i>. EC numbers are given on each enzymatic reaction. Light blue font indicates concentrations (per pmol 10<sup>10</sup> cells) of metabolites. Metabolites that were not targets of the metabolomic analysis were given neither concentrations nor N.D. N.D.; not detected. Pi, phosphate; PPi, pyrophosphate; Glc, α-D-glucose; αG6P, α-D-glucose-6-phosphate; G1P, α-D-glucose-1-phosphate; βG6P, β-D-glucose-1-phosphate; F6P, β-D-fructose-6-phophate; FBP, β-D-fructose-1,6-bisphosphate; GAP, glyceraldehyde-3-phosphate; GBP, glycerate-1,3-bisphosphate; G3P, glycerate-3-phosphate; G2P, glycerate-2-phosphate; PEP, phosphoenolpyruvate; E4P, erythrose-4-phosphate; acetyl-P, acetyl phosphate; DHAP, dihydroxyacetone phosphate; X5P, D-xylulose-5-phosphate; SBP, D-sedoheptulose-1,7-bisphosphate; S7P, D-sedoheptulose-7-phosphate; RuBP, ribulose-1,5-bisphosphate; Ru5P, ribulose-5-phophate; R5P, D-ribose-5-phosphate.</p
Electron micrographs of the new isolate strain Hiromi 1.
<p>A transmission electron micrograph of a negatively stained cell (A) and a thin section cell (B) grown under the chemolithoautotrophic condition. White and black arrows indicate intracellular particle and outer membrane, respectively. Scanning micrographs of cells adhering on elemental sulfur by pilus to biofilm (C, D). Cells attached on biofilm formation and pili structures were shown by large and small black arrows. Other cells grew under the polysaccharide-like substances. Bars, 0.2 µm (A), 0.5 µm (B) and 1 µm (C, D).</p
A SSU rRNA gene phylogenetic tree of chemolithoautotrophic and methanotrophic <i>Gammaproteobacteria</i> including strain Hiromi 1 constructed by the neighbor-joining method using 1253 identical positions.
<p>Bootstrap values higher than 50% are presented. GenBank/EMBL/DDBJ accession numbers are given in parentheses. Bar indicates 1 substitutions per 100 nucleotides.</p
Isolation, characterization and genomic features of sulfur-oxidizing Gammaproteobateria belongs to a chemoautotrophic endosymbionts lineage
http://www.godac.jamstec.go.jp/darwin/cruise/natsushima/nt07-11/