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| Status |
Public on Jun 01, 2012 |
| Title |
Transcriptional profiling of equine chondrocyte de-differentiation in culture |
| Organism |
Equus caballus |
| Experiment type |
Expression profiling by array
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| Summary |
Monolayer cultures of primary chondrocytes undergo morphological changes that have been broadly characterized as de-differentiation. Transcriptional profiling was used to evaluate changes in gene expression during this process. Articular cartilage was harvested from a single Standardbred horse; the chondrocytes were isolated and then maintained in monolayer culture for up to 14 days. Chondrocytes were collected on Day 1 and Day 14 of culture and snap frozen in liquid nitrogen. Total RNA was isolated, subjected to one round of linear RNA amplification, and then applied to an equine-specific cDNA microarray composed of 9322 elements. Three biological replicates were analyzed at each time point using a dye-swap experimental design. One hundred six transcripts were present at levels greater than or equal to five-fold higher in Day 1 chondrocytes relative to the cells collected at Day 14. Conversely, 68 transcripts were present at levels greater than or equal to five-fold higher in Day 14 chondrocytes. Northern blot hybridization validated the microarray data by confirming the down-regulation of steady state mRNA levels for type II procollagen and aggrecan core protein and the up-regulation of type I procollagen. Gene-by-gene and cluster analyses will be used to compare the process of “de-differentiation” in primary culture to the developmental stages of normal chondrocyte differentiation and to the fibrocartilage repair tissue that forms in osteoarthritic lesions.
Keywords: articular cartilage, de-differentiation, horse, cDNA microarray
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| Overall design |
This is a direct comparison between one and fourteen day cultured chondrocyte transcriptomes. Three sets of technical replicates were compared. For each comparison and its dye-swap, one Day 1 technical replicate was randomly compared to one Day 14 technical replicate.
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| Contributor(s) |
Mienaltowski MJ, Stromberg AJ, MacLeod JN |
| Citation missing |
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| Submission date |
Dec 31, 2008 |
| Last update date |
Jun 01, 2012 |
| Contact name |
Michael J. Mienaltowski |
| E-mail(s) |
M.Mski@uky.edu, mmienalt@health.usf.edu
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| Phone |
8592574757
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| Organization name |
University of Kentucky
|
| Department |
Veterinary Science
|
| Lab |
MacLeod Laboratory
|
| Street address |
108 Gluck Equine Research Center
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| City |
Lexington |
| State/province |
KY |
| ZIP/Postal code |
40546 |
| Country |
USA |
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| Platforms (1) |
| GPL6423 |
MacLeod custom equine cartilage cDNA microarray |
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| Samples (3) |
| GSM356962 |
Day 14 vs. Day 1 Chondrocyte Comparison A |
| GSM356963 |
Day 14 vs. Day 1 Chondrocyte Comparison B |
| GSM356964 |
Day 14 vs. Day 1 Chondrocyte Comparison C |
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| Relations |
| BioProject |
PRJNA111209 |
