Expression data from rat spinal cord injury and mesenchymal stromal cells (MSC) or olfactory ensheathing cells (OEC) transplantation
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ABSTRACT: We analyzed the changes in the spinal cord transcriptome after a spinal cord contusion injury and MSC or OEC transplantation. The cells were injected immediately or 7 days after the injury. The mRNA of the spinal cord injured segment was extracted and analyzed by microarray at 2 and 7 days after cell grafting.
Project description:We analyzed the changes in the spinal cord transcriptome after a spinal cord contusion injury and MSC or OEC transplantation. The cells were injected immediately or 7 days after the injury. The mRNA of the spinal cord injured segment was extracted and analyzed by microarray at 2 and 7 days after cell grafting. 52 total samples were analyzed in 13 different groups. Each group include 4 samples and each one were analyzed as a biological replica. The intact animals were used as control of injury. The vehicle (VHC) groups were used as control of transplantation procedure. The MSC or OEC graft were injected at the day of injury (acute graft) or seven days after injury (delayed graft). The samples from engrafted animals were obtained at 2 or 7 days after cell transplantation. To determine the effects of MSC or OEC transplantation, the expression value of each engrafted sample were compared with correspondent VHC group.
Project description:We profiled spinal cord tissue at the site of a moderate contusion injury at the level of the thoracic spinal cord We examined several timepoints following injury, including sham and days 1,3 and 7 following injury and compared differential expression of genes within a genotype and across genotypes (trkB.T1KO/trkB.T1WT) at each timepoint. Tissue was profiled at baseline (sham) condition and then 1, 3 and 7 days after thoracic moderate contusion injury
Project description:Excerpt from a larger study which characterized the transcriptional effects of a spinal cord contusion injury in rats. This is the data from the almost chronic contusion state (35 days) at the injury site (Thoracic 8) - where we saw significant changes in several areas, including cholesterol metabolism genes. Other spinal cord areas (rostral, caudal) and time-points (3 hours, 24 hours, 7 days and 35 days) were analyzed as well and are discussed in our paper and at www.crpf.org/microarray. Keywords = Spinal Cord Injury Keywords = chronic Keywords = thoracic Keywords = cholesterol Keywords: repeat sample
Project description:Excerpt from a larger study which characterized the transcriptional effects of a spinal cord contusion injury in rats. This is the data from the almost chronic contusion state (35 days) at the injury site (Thoracic 8) - where we saw significant changes in several areas, including cholesterol metabolism genes. Other spinal cord areas (rostral, caudal) and time-points (3 hours, 24 hours, 7 days and 35 days) were analyzed as well and are discussed in our paper and at www.crpf.org/microarray.
Project description:Identification of temporal variations in miRNA expression after spinal cord injury caused by thoracic (T8) moderate (200 Kdynes) contusion. Expression changes were analyzed 1, 3 and 7 days after injury and compared to expression of control (untreated) and sham (laminectony but no contusion) individuals.
Project description:We profiled spinal cord tissue at the site of a moderate contusion injury at the level of the thoracic spinal cord We examined several timepoints following injury, including sham and days 1,3 and 7 following injury and compared differential expression of genes within a genotype and across genotypes (trkB.T1KO/trkB.T1WT) at each timepoint.
Project description:Comparison of three populations of OECs derived from the ONL of rats. The three populations were cultured employing the same culture medium but were maintained in vitro for a different amount of time (number of population doublings or PDs). OEC Ep cells were maintained for less than one week in vitro (less than 3 PDs); OEC Lp were maintained for over a year (more than 50 PDs) and TEG3 OEC cell line has been serially passed in culture for long-periods of time after genetic immortalization. Experiment Overall Design: RNA sample preparation and experimental design. Total RNA was isolated with TRIZOL (Invitrogen, Gibco-BRL) from primary OEC cultures (OEC Ep and Lp) and the OEC cell line (TEG3). Cultured cells were harvested at confluency with a rubber scrapper, resuspended in the TRIZOL reagent and processed following the manufacturersâ?? instructions. Two RNA pools were obtained from each OEC population (OEC Ep, OEC Lp and TEG3 cells). Each RNA pool was purified from three individual cultures (three 100mm dishes) and in the case of OEC Ep, each culture was derived from one rat. The total RNA was resuspended in DEPC (Sigma) water and stored at -70ºC prior to use. Total RNA concentration and purity were determined using Agilent2100 Bioanalizer. Experiment Overall Design: Microarray analysis. Total RNA pools were cleaned using Rneasy Kit from Qiagen. Biotinylated cRNA probes were generated from each RNA pool, fragmented, and applied to two different sets of Rat Genome Affymetrix GeneChip U34A Array. Affymetrix software (Microarray Suite version 5.0) was used to filter inaccurately represented probe sets.
Project description:Identification of temporal variations in miRNA expression after spinal cord injury caused by thoracic (T8) moderate (200 Kdynes) contusion. Expression changes were analyzed 1, 3 and 7 days after injury and compared to expression of control (untreated) and sham (laminectony but no contusion) individuals. Included groups: control (untreated), 1 day after lesion, 3 days after lesion, 7 days after lesion, 1 days after SHAM cirugy, 3 days after SHAM cirugy, and 7 days after SHAM cirugy. Each of these 7 groups included 5 biological replicates.
Project description:Olfactory ensheathing cells (OECs) are the only glial cells that support the olfactory sensory neurons which undergo adult neurogenesis and continually project their axons to glomeruli in the olfactory bulbs. We used single cell RNA sequencing to study the gene expression programs of OECs and to determine the diversity of purified OECs previously shown to promote spinal cord injury repair. Our analyses revealed five subtypes of OECs, each expressing unique marker genes and pathways indicative of progenitor, axonal regeneration, migration, or microglia-like functions. As expected, we found substantial overlap of OEC genes with those of Schwann cells, but also with astrocytes, oligodendrocytes and microglia. We experimentally confirmed the classic marker genes of the OEC subtypes and provide evidence that Reelin and Connective Tissue Growth Factors are secreted by multiple OEC subtypes. Our results support that adult OECs are a hybrid glia including some with progenitor characteristics and that they likely carry out diverse functions related to injury repair and axonal regeneration.
Project description:Traumatic spinal cord injury (SCI) initiates a complex series of pathophysiological secondary responses that lead to tissue loss and functional deficits.This study represents a comprehensive database of temporal changes in gene expression that underlie the secondary injury response that occurs in a well-defined mouse model of contusion injury. Experiment Overall Design: Global changes in gene expression that contribute to secondary injury in a C57BL6 mouse model of contusion injury were evaluated using Affymetrix Mouse Genome 430 2.0 arrays.Mice were subjected to a moderate injury at the T8 spinal segment under isoflurane anesthesia.Sections of cord (0.4 cm in length) were analyzed from the site of impact and from the immediately adjacent rostral and caudal regions at 0.5, 4, 24, 72 h and 7 and 28 days after injury (n=3/per group), sham-injury (n=2 per group), or from naive mice (n=2). Four mice were pooled for each individual n, for a total of 12 mice for each injury time point.