Dataset Information

Bacteria and Fungi sampled from a Naked Mole Rat (Heterocephalus glaber)

ABSTRACT: Microbial isolates sourced from H. glaber skin, small intestine, and large intestine. Isolates were grown on solid ISP-2 agar. Individual colonies underwent lysis in pure TFA before extraction in ACN:H2O solution. The resulting extract was spotted on the target plate in a 1:1 ratio with CHCA matrix. Originally collected to be analyzed through the IDBac workflow. Metabolite (reflectron, 200-2000 Da) and protein (linear, 2000-20,000 Da) spectra were collected in triplicate for each isolate. Technical replicates are sequential left to right within a row, (A1, A2, and A3 are all replicates). CHCA matrix. Matrix blank for each run can be found in the bottom right-most target. Only calibrants and the matrix blank are on the final row (letter) of each plate. Additional information on the project can be found in the thesis published in the UIC INDIGO thesis database under the title "Using Mass Spectrometry to Quantitate and Analyze Bioactive Small Molecules", author last name "Grim".

INSTRUMENT(S): autoflex speed LRF (Bruker Daltonics flex series)

ORGANISM(S): Heterocephalus Glaber (ncbitaxon:10181) Environmental Samples <bacillariophyta> (ncbitaxon:33858)

SUBMITTER: Laura Sanchez

PROVIDER: MSV000085215 | MassIVE | Thu Apr 02 02:00:00 BST 2020

REPOSITORIES: MassIVE

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Project description:Human pluripotent stem cells (hPSCs) are a promising source of cells for applications in regenerative medicine. Directed differentiation of hPSCs into specialized cells such as spinal motoneurons or midbrain dopamine (DA) neurons has been achieved. However the effective use of hPSCs for cell therapy has lagged far behind. While mouse PSC-derived DA neurons have shown efficacy in models of Parkinson’s disease, DA neurons derived from human PSCs generally display poor in vivo performance. There are also considerable safety concerns for hPSCs related to their potential for teratoma formation or neural overgrowth. Here we present a novel floor plate-based strategy for the derivation of human DA neurons that efficiently engraft, suggesting that past failures were due to incomplete specification rather than a specific vulnerability of the cells. Midbrain floor plate precursors are derived from hPSCs in days following exposure to small molecule activators of sonic hedgehog (SHH) and canonical WNT signaling. Engraftable midbrain DA neurons are obtained by day 25 and can be maintained in vitro for several months. Extensive in vitro molecular profiling, biochemical and electrophysiological data define developmental progression and confirm identity of hPSC-derived midbrain DA neurons. In vivo survival and function is demonstrated in PD animal models in three host species. Long-term engraftment in 6-OHDA-lesioned mouse and rats demonstrates robust survival of midbrain DA neurons, complete restoration of amphetamine-induced rotation behavior and improvements in tests of forelimb use and akinesia. Finally, scalability is demonstrated by transplantation into Parkinsonian monkeys. Excellent DA neuron survival, function and lack of neural overgrowth in the three animal models tested indicate considerable promise for the development of cell based therapies in PD. Differentiated hESC with three conditions (LSB, LSB/S/F8, LSB/S/F8/CHIR) were subjected to RNA extraction in specific timepoint (day 0, 1, 3, 5, 7, 11, 13, 25) and hybridization on Illumina microarrays. Each sample has 3 or 4 biological repeats. Based on previous study* of dual SMAD inhibition neural induction, we developed new midbrain dopamine neuron protocol. It depends on time specific treatment of below factors (LSB/S/F8/CHIR): L (LDN193189 (BMP inhibitor) , day 0-11), SB (SB431542 (TGF-b signal inhibitor), day 0-5), S (SHH + Purmorphamine (Smo agonist), day 1-7), F8 (FGF8, day 1-7) and CHIR (CHIR99021 (GSK3b inhibitor), day 3-13) LSB and LSB/S/F8 are limited control conditions of dual SMAD only (LSB) or traditional patterning with Sonic and FGF (LSB/S/F8) *Chambers,S.M. et al. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat. Biotechnol. 27, 275-280 (2009).

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Bacteria and Fungi sampled from a Naked Mole Rat (Heterocephalus glaber)

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