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Resuscitation from Near Cell Death Unleashes Developmental Cues that Enhance Tissue Repair and Regeneration

ABSTRACT: Resuscitation from near-cell death conditions is an intriguing but not well-understood cell biology phenomenon. First, we conclusively demonstrate that cells could revive from the early cell death phase evoked by LLOMe and other lysosomotropic agents. The resuscitation phase was found to be a highly orchestrated cellular and molecular process, where the fragmented cell organelles of the death phase fully renewed themselves within a few hours. Remarkably, the initiation of the resuscitation phase was accompanied by the upregulation of pathways of embryonic development, tissue repair, regeneration, and stemness. This was followed by the induction of networks of genes of vital cellular and metabolic pathways. Prompted by in vitro results, we tested whether LLOMe could affect tissue repair, regeneration, and stemness in vivo. Indeed, LLOMe dramatically induced repair in skin wound and corneal alkali burns mouse models. Also, it stimulated the production of hematopoietic stem/progenitor cells in the Drosophila melanogaster, enhanced tadpole tail regeneration in the frog, and promoted axon regeneration in the Caenorhabditis elegans. Fascinatingly, LLOMe treatment of frog tadpoles resulted in accelerated developmental growth. Among others, NF-ĸB signaling was found to be most critical for the revival of cells as well as the regeneration process. Thus, for the first time, we demonstrate that resuscitation from near-cell death conditions is an elegantly programmed process and that this process could be harnessed for therapeutic advances in regenerative medicine.

ORGANISM(S): Mus musculus

PROVIDER: GSE285446 | GEO | 2025/07/01

REPOSITORIES: GEO

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Project description:Objective of the study is to find out the differentially regulated genes of Mycobacterium BCG strain in extended stationary phase comparison to log phase growth and resuscitation phase. Mycobacterium BCG culture was grown in Sauton medium at 37o C without shaking. Cells at A600 0.6-0.8 were harvested as log phase culture. The cells harvested after 5 months incubation at 37oC without shaking became non culturable and were harvested as extended stationary phase cells. The extended stationary phase cells were treated with resuscitation promoting factor (Rpf) from Micrococcus luteus and harvested after 8 days. Cells of this stage were treated as resuscitation phase cells. Gene expression profiling was carried out using Agilent microarray platform. Keywords: Extended stationary phase, Log phase growth and Resuscitation phase. â¢ Organism: Mycobacterium tuberculosis BCG â¢ Slides: Agilentâs Mycobacterium tuberculosis custom array 4x44k (G2514F) AMADID No: 016013 â¢ Labeling kit: Ambion Message Amp II Bacteria a RNA Amplification Kit Cat# 1790 â¢ Labeling Method: T7 promoter based-linear amplification to generate labeled complementary RNA â¢ Total RNA and cRNA Purification Kit: Qiagenâs RNeasy minikit Cat#74104 â¢ Hybridization Kit: Agilentâs In situ Hybridzation kit 5184-3568 â¢ RNA quality was checked using Bioanalyzer. Hybridization protocol 825ng each of labeled control cRNA was mixed with 825ng of treated labelled cRNA, mixed with appropriate amount of blocking buffer and hybridization buffer (Agilent Technologies). The hybridization mix was applied on to the backings and hybridized to custom designed 60mer Mycobacterium tuberculosis microarray using sure hyb chambers at 65degC for 17 hours. Slides were washed with gene expression wash buffer 1 and 2 (Agilent Technologies) followed by Acetonitrile. Scan protocol Laser detection of Cyanine 3 and Cyanine 5 fluorescence is performed using a confocal scanning instrument containing two tuned lasers, which excite Cyanine dyes at the appropriate wavelengths. Description Slides were scanned at 5 micron resolution using Agilent scanner.Automated feature extraction was done using Agilentâs Feature Extraction Software. Data processing Analysis of feature extracted data was done using Agilentâs GeneSpring GX V 7.3.1 software. Pre-processing of the replicate experiment data was done to flag the outliers. Normalization of the data was done using per spot per chip intensity dependant lowess normalization and dye swap experiment was data transformed. Genes with log2 ratio of 1.29 and above in replicate experiments were considered as up regulated and 0.81 and below was considered down regulated. p-value was calculated by GeneSpring GX for each gene on the basis of replicate probes to indicate statistical significance. Biological significance of differentials were analyzed using Genotypics Biointerpreter - Web based tool for biological interpretation of given genelist.

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Resuscitation from Near Cell Death Unleashes Developmental Cues that Enhance Tissue Repair and Regeneration

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