Tissue processing doesn't need to be complex and time consuming. STEMprep™ takes the hassle out of sample prep with a sleek, customizable interface and built-in temperature control to keep your samples cool and your workflow smooth. Less time prepping, more time generating data that you trust—that’s the power of STEMprep™.
The STEMprep™ Tissue Dissociator enables efficient processing of a wide range of tissues, including mouse brain, liver, lung, spleen, and tumors. In this workflow, tissue samples are placed into a STEMprep™ Sample Tube and processed using the STEMprep™ Tissue Dissociator in combination with the appropriate STEMprep™ Tissue Dissociation Kit. Depending on the selected protocol-dissociation or homogenization-the resulting output is either a single-cell suspension or a sample suitable for nucleic acid isolation. STEMprep™-processed cells are compatible with downstream applications such as cell separation, cell culture, flow cytometry, and other assays.
Figure 2. STEMprep™ Cold Temperature Hold Feature Preserves Mouse Lung and Spleen Cell Viability
Mouse lung and spleen tissues were dissociated using the STEMprep™ Mouse Lung and Spleen Dissociation Kits, respectively. After the protocol run was completed, the samples were held on the STEMprep™ Tissue Dissociator for up to 24 hours at 4°C, 22°C, and 37°C. Viability of total nucleated (A) lung and (B) spleen cells. Data are presented as mean ± SD (n = 3).
Figure 3. RNA Yields from Tissues Homogenized Using the STEMprep™ Tissue Dissociator Are Comparable to Other Common Homogenization Methods.
Mouse (A) brain, (B) lung, or (C) liver tissues were homogenized using the STEMprep™ Tissue Dissociator or an alternative automated system. RNA was subsequently extracted using the EasySep™ Total Nucleic Acid Extraction Kit (with DNase I treatment). The yield of RNA per mg of tissue was assessed using the Qubit Flex Instrument and Qubit RNA Broad Range Assay kit. Data are presented as mean ± SD (n = 8).
Figure 4. STEMprep™ Direct Tissue Homogenization Yields High-Quality RNA
RNA integrity number (RIN) of RNA extracted from mouse (A) brain, (B) lung, or (C) liver tissues. Tissues were homogenized using the STEMprep™ Tissue Dissociator or an alternative automated system. RNA was subsequently extracted from the resulting homogenates by EasySep™ Total Nucleic Acid Extraction Kit (with DNase I treatment). The quality of the extracted RNA was assessed using the 2100 Bioanalyzer Instrument and Agilent RNA 6000 Nano Kit. Data are presented as mean ± SD (n = 8).
Figure 5. STEMprep™ Template Protocol Information
Step 1 (Prime): A brief slow spin during pre-heat.
Steps 2 and 3 (Cut): Steps 2 and 3 are looped with a repeat count of 2. A reverse spin (-40 RPM) is applied to step 3 to ensure free movement of tissue.
Note: Reduce speed for soft tissues. Increase speed or number of loops for hard tissues.
Step 4 (Incubate): Slow spin to agitate tissue during incubation with enzyme cocktail.
Step 5 (Dissociate): High-speed spin to generate single cells.
Microbial dysbiosis sculpts a systemic ILC3/IL-17 axis governing lung inflammatory responses A Kabil et al.
Mucosal Immunology 2025 July
Abstract
Advancements in vaccination and sanitation have significantly reduced the prevalence and burden of infectious diseases; however,these benefits have coincided with a marked rise in autoimmune and allergic disorders. Recent studies have investigated these linked trends through the lens of host-microbiome alterations,proposing these shifts as a potential explanatory mechanism. Previously,we demonstrated that vancomycin-induced depletion of short-chain fatty acid (SCFA)-producing bacteria results in hyperactivation of ILC2s and exacerbated allergic responses. Here we investigate the effects of low-dose streptomycin on innate and adaptive immune cell populations and their activation states. Although streptomycin-treated mice exhibit normal allergic responses,they display heightened susceptibility to Th1/Th17-mediated disease,specifically hypersensitivity pneumonitis (HP). This is characterized by a two-fold increase in ILC3s and Th17 cells in the lungs,alongside activation of antigen-presenting cells (APCs) at steady state-an effect that is further amplified upon exposure to HP-inducing agents. Shotgun metagenomic analysis revealed that streptomycin-induced dysbiosis reduces microbial diversity,depletes bile acid-metabolizing bacteria,and enriches for metabolic pathways involved in branched-chain amino acid biosynthesis,including leucine-a known activator of mTORC1. Strikingly,administration of the secondary bile acid metabolite isolithocholic acid (an inverse agonist of RORγt),or an IL-23 neutralizing antibody,reverses the enhanced susceptibility to HP. Inhibition of mTORC1 significantly reduced Th17/ILC3 responses and histopathology. Our findings underscore microbial equilibrium as a key determinant of susceptibility to HP and uncover a positive feedback loop between IL and 23-producing APCs and ILC3/Th17 cells that mechanistically links dysbiosis to sustained type 3 inflammation,and we identify a simple,actionable means of intervention.
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