Release of tissue inhibitor of metalloproteinase-2 from alginate microcapsule encapsulating genetically engineered cells.
BACKGROUND
In this study, 293T cells were genetically engineered to secrete tissue inhibitor of metalloproteinase-2 (TIMP2) and encapsulated into alginate microcapsules to continuously release TIMP2 protein.


METHODS
The anti-invasive potential of the microcapsules was studied in vitro using brain tumor cells. The TIMP2 gene was transfected to 293T cells, and genetically engineered 293TIMP2 cells were encapsulated into alginate microcapsules. Release of TIMP2 protein was detected with Western blot analysis and the anti-invasive potential against U87MG cells was tested using gelatin zymography and a Matrigel assay.


RESULTS
Cell viability within the alginate microcapsules was maintained at a cell density of 5 × 10(6). Because polycationic polymers are helpful for maintaining the mechanical strength of microcapsules with good cell viability, the alginate microcapsules were reinforced with chitosan (0.1% w/v). Expression of TIMP2 protein in cell lysates and secretion of TIMP2 into the conditioned medium was confirmed by Western blot analysis. Alginate microcapsules encapsulating 293TIMP2 cells released TIMP2 protein into the medium efficiently, where the TIMP2 protein participated in degradation of the matrix metalloproteinase-2 enzyme and inhibited invasion of U87MG cells.


CONCLUSIONS
Alginate microcapsules encapsulating 293TIMP2 cells are promising candidates for anti-invasive treatment of glioma.
Department of Neurosurgery, Chonnam National University Hwasun Hospital and Medical School, Jeollanam-do, Korea.
Yeon Seong
YS
Kim
Young-Ii
YI
Jeong
Shu-Guang
SG
Jin
Jian
J
Pei
Min
M
Wen
In-Young
IY
Kim
Kyung-Sub
KS
Moon
Tae-Young
TY
Jung
Hyang-Hwa
HH
Ryu
Shin
S
Jung
D000464:Alginates / Q000737:chemistry*; D002214:Capsules / Q000737:chemistry* / Q000378:metabolism; D060846:Cell Engineering; D005818:Genetic Engineering / Q000379:methods*; D020723:Glucuronic Acid / Q000737:chemistry; D057809:HEK293 Cells; D006603:Hexuronic Acids / Q000737:chemistry; D006801:Humans; D010316:Particle Size; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics* / Q000378:metabolism*; D014162:Transfection
D000464:Alginates; D002214:Capsules; D006603:Hexuronic Acids; C560823:TIMP2 protein, human; D019716:Tissue Inhibitor of Metalloproteinase-2; D020723:Glucuronic Acid
293T cells; alginate microcapsule; therapeutic protein; tissue inhibitor of metalloproteinase-2