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The expression of TIMPs in cryo-preserved and freeze-dried amniotic membrane
Cited 23 time in
Web of Science
Cited 26 time in Scopus
- Authors
- Issue Date
- 2007-09-14
- Publisher
- Taylor & Francis
- Citation
- Curr Eye Res. 2007 Jul-Aug;32(7-8):611-6.
- Keywords
- Amnion/*metabolism ; Blotting, Western ; Epithelial Cells/metabolism ; Freeze Drying ; Gene Expression/physiology ; Humans ; Immunoenzyme Techniques ; RNA, Messenger/metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Stromal Cells/metabolism ; Tissue Inhibitor of Metalloproteinase-1/*genetics/metabolism ; Tissue Inhibitor of Metalloproteinase-2/*genetics/metabolism ; Cryopreservation ; Tissue Preservation
- Abstract
- PURPOSE: To investigate the change of tissue inhibitor of metalloproteinase (TIMP) in cryopreserved amniotic membranes (AM) according to preservation time, and to evaluate the expression of TIMP in freeze-dried AM. METHODS: Cryopreserved or fresh AMs were incubated in dispase II for two hours at 37 degrees C and their epithelial cells were scraped with a cell scraper. Remaining stromal AM was minced and frozen in liquid nitrogen, and then treated with 0.1% diethyl pyrocarbonate. The mRNA levels of TIMP-1 and -2 were determined by reverse transcription-polymerase chain reaction (RT-PCR) in epithelial and stromal cells of fresh AM, AMs cryopreserved for 6 and 12 months, and freeze dried AM, respectively. Western blot analysis and immunohistochemical staining were performed to assess the expression of TIMP-1 in fresh, cryopreserved, and freeze dried AMs. RESULTS: RT-PCR revealed that mRNAs of TIMP-1 and -2 were expressed in the amniotic epithelial cells of both fresh and cryopreserved AMs, while the stromal cells of fresh or cryopreseved AMs and freeze-dried AM showed higher expression of TIMP-1 than TIMP-2 mRNA. On Western blot analysis, the level of TIMP-1 was more in fresh AMs than in cryopreserved or freeze-dried AM, but it was not statistically significant. CONCLUSION: TIMP-1 was expressed in cryopreserved AMs until 12 months, and the amount of expression was comparable to that in fresh AMs.
- ISSN
- 0271-3683 (Print)
- Language
- English
- URI
- http://www.informaworld.com/smpp/content~db=all?content=10.1080/02713680701459441
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17852184
https://hdl.handle.net/10371/15677
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