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Changwon Kee, Seongsoo Sohn, Jeong-Min Hwang; Stromelysin Gene Transfer into Cultured Human Trabecular Cells and Rat Trabecular Meshwork In Vivo. Invest. Ophthalmol. Vis. Sci. 2001;42(12):2856-2860. doi: https://doi.org/.
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purpose. To determine whether stromelysin gene can be introduced into
and expressed in the cultured human trabecular cells as well as in the
rat eye in vivo through means of a recombinant replication-deficient
methods. Stromelysin cDNA was obtained by reverse transcription-polymerase chain
reaction with mRNA extracted from the cultured human trabecular cells
after induction with interleukin 1α. Adenovirus vector that contains
stromelysin cDNA was constructed by cotransfection of pJM17 and
pΔA.CMV-str into the 293 cells. The expression of stromelysin
in the cultured human trabecular cells was assayed by Western blot and
zymography. The expression of stromelysin in the trabecular meshwork of
the rat eyes was detected by in situ hybridization and
results. The constructed adenovirus vector contained stromelysin cDNA, but no E1
region. Western blot and zymogram revealed that the stromelysin could
be expressed and that it possessed enzymatic activity in cultured human
trabecular cells. In situ hybridization and immunostaining of the
stromelysin showed that the complete form of stromelysin was expressed
in the trabecular meshwork, the iris, and the uveoscleral outflow
pathway of the rat eye.
conclusions. Stromelysin, a functional gene, can be transferred in vivo into rat
eyes and in vitro into cultured human trabecular cells using a
replication-deficient adenovirus vector. This shows the possibility of
gene therapy in glaucoma.
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