FUNCTIONAL HUMAN SPINAL CORD TISSUE & CELLS
AnaBios recovers human spinal cord tissue and cells in a highly viable state in order to probe the effects of compounds on spinal cord neurons. We have developed highly specialized recovery techniques to maintain tissue and cell viability such that the tissues can be sliced by vibratome and maintained in culture for weeks (organotypic culture).
Researchers interested in the transcriptomic or proteomic signature of human spinal cord can acquire flash frozen or fixed human spinal cord samples at specified vertebrate levels with confidence that the tissue quality remains at the same level as human spinal cord used for organotypic culture.
HUMAN SPINAL CORD TISSUE & CELLS RIN SCORES
Maintenance of viable human spinal cord tissue samples and cells is critical to generating high-quality transcriptomic or proteomic data. Initiation of ischemic insult associated with the surgical recovery of human organs can trigger necrotic and apoptotic pathways that degrade RNA and proteins.
AnaBios checks the quality of RNA in recovered human tissue samples from each donor by extracting the RNA and assessing the amount of degradation through the RNA Integrity Number (RIN).
An Agilent Bioanalyzer, the agreed gold standard for RNA integrity determination, is used to create RIN score. The average RIN score for human spinal cord tissue recovered by AnaBios is greater than 9, while the threshold for high quality RNA for use in expression analysis is 7.
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The left panel of Figure 1 (see below) shows calcium-sensitive fluo-8 fluorescence from neurons in a spinal cord slice. 30μM bicuculine was added, causing spontaneous calcium oscillations as exhibited in the right panel of nine identified neurons (arrows in left panel).
Figure 1: Calcium-sensitive Fluorescence from Spinal Cord Slice
In a separate experiment, spontaneous calcium activity was recorded in a spinal cord slice. Figure 2 (see below) shows a raster plot with sticks identifying the time location of the peaks of spontaneous calcium transients. 1μM TTX was added at 20 minutes which caused an inhibition of the spontaneous transient.
Figure 2: Raster Plot from Human Spinal Cord Slice Activity
CGRP IMMUNOSTAINING OF SPINAL CORD TISSUE
Figure 1: Human Spinal Cord Tissue Slice
The spinal cord section in Figure 1 (above), is magnified in Figure 2 (below), with a subsection showing detailed spinal lesion. (Images courtesy of Michael Iadarola, Matthew Sapio)
Figure 2: Magnified Subsection of Spinal Section from Spinal Cord
HUMAN SPINAL CORD RESEARCH
In the short video above, Fluo-8 loaded cells in the human dorsal horn can be seen. In the image above is a human spinal cord slice cut on a Leica vibratome and stained with TTC, which is a colometric marker for active mitochondria and a marker of viable neurons. In this section, TTC marks the spinal cord’s white matter. (Photo credits: Tamara Cotta, AnaBios)
SPINAL CORD TISSUE APPLICATIONS
Preserved human spinal cord tissue samples can be used to support proteomic and transcriptomic studies of the human spinal cord. Spinal cord samples are flash frozen, preserved in RNAlater or fixed by formalin- based solutions. These human tissue samples can be used for bulk extraction of RNA or for proteomic and expression analysis; or they can be processed for RNAscope or immunohistochemistry.
PAIN RESEARCH WEBINARS SPONSORED BY ANABIOS
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