The expression of TIMPs in cryo-preserved and freeze-dried amniotic membrane

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Koh, Jae Woong; Shin, Young Joo; Oh, Joo Youn; Kim, Mee Kum; Ko, Jung Hwa; Hwang, Jeong Min; Wee, Won Ryang; Lee, Jin Hak
Issue Date
Taylor & Francis
Curr Eye Res. 2007 Jul-Aug;32(7-8):611-6.
Amnion/*metabolismBlotting, Western*CryopreservationEpithelial Cells/metabolismFreeze DryingGene Expression/physiologyHumansImmunoenzyme TechniquesRNA, Messenger/metabolismReverse Transcriptase Polymerase Chain ReactionStromal Cells/metabolismTissue Inhibitor of Metalloproteinase-1/*genetics/metabolismTissue Inhibitor of Metalloproteinase-2/*genetics/metabolism*Tissue Preservation
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.
0271-3683 (Print)
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College of Medicine/School of Medicine (의과대학/대학원)Ophthalmology (안과학전공)Journal Papers (저널논문_안과학전공)
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