Full Text:   <1336>

CLC number: R73

On-line Access: 

Received: 2005-04-28

Revision Accepted: 2005-05-24

Crosschecked: 0000-00-00

Cited: 1

Clicked: 4133

Citations:  Bibtex RefMan EndNote GB/T7714

-   Go to

Article info.
1. Reference List
Open peer comments

Journal of Zhejiang University SCIENCE B 2005 Vol.6 No.9 P.859~864

10.1631/jzus.2005.B0859


Preparation and in vitro studies of microencapsulated cells releasing human tissue inhibitor of metalloproteinase-2


Author(s):  JIANG Qiang, ZHANG Su-zhan, PENG Jia-ping, WANG Xu-lin

Affiliation(s):  Cancer Institute, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China

Corresponding email(s):   jiangtiger@zju.edu.cn, zhangscy@tom.com

Key Words:  Microencapsulation, Recombinant cells, Human tissue inhibitor of metalloproteinase-2, Cell culture


JIANG Qiang, ZHANG Su-zhan, PENG Jia-ping, WANG Xu-lin. Preparation and in vitro studies of microencapsulated cells releasing human tissue inhibitor of metalloproteinase-2[J]. Journal of Zhejiang University Science B, 2005, 6(9): 859~864.

@article{title="Preparation and in vitro studies of microencapsulated cells releasing human tissue inhibitor of metalloproteinase-2",
author="JIANG Qiang, ZHANG Su-zhan, PENG Jia-ping, WANG Xu-lin",
journal="Journal of Zhejiang University Science B",
volume="6",
number="9",
pages="859~864",
year="2005",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.2005.B0859"
}

%0 Journal Article
%T Preparation and in vitro studies of microencapsulated cells releasing human tissue inhibitor of metalloproteinase-2
%A JIANG Qiang
%A ZHANG Su-zhan
%A PENG Jia-ping
%A WANG Xu-lin
%J Journal of Zhejiang University SCIENCE B
%V 6
%N 9
%P 859~864
%@ 1673-1581
%D 2005
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2005.B0859

TY - JOUR
T1 - Preparation and in vitro studies of microencapsulated cells releasing human tissue inhibitor of metalloproteinase-2
A1 - JIANG Qiang
A1 - ZHANG Su-zhan
A1 - PENG Jia-ping
A1 - WANG Xu-lin
J0 - Journal of Zhejiang University Science B
VL - 6
IS - 9
SP - 859
EP - 864
%@ 1673-1581
Y1 - 2005
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.2005.B0859


Abstract: 
Objective: To prepare microencapsulated cells releasing human tissue inhibitor of metalloproteinase-2 (TIMP-2), and investigate their biological characteristics in vitro. Methods: Chinese hamster ovary (CHO) cells were stably transfected with a human TIMP-2 expression vector, encapsulated in barium alginate microcapsules and cultured in vitro. Morphological appearance of the microcapsules was observed under a light microscope. Cell viability was assessed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Enzyme linked immunosorbent assay (ELISA) and reverse zymography were used to confirm the release of biologically active TIMP-2 from the microcapsules. Cryopreservation study of the microencapsulated cells was carried out using dimethyl sulfoxide (DMSO) as preservative agent. Results: The microcapsules appeared like a sphere with diameter of 300~600 μm. The surface of the capsule wall was clearly smooth. The microencapsulated cells survived well and kept proliferating over the 6 weeks observed. No significant difference in TIMP-2 secretion was found between encapsulated and unencapsulated cells. Reverse zymography confirmed the bioactivity of MMP (matrix metalloproteinase) inhibition of TIMP-2. The cryopreservation process did not damage the microcapsule morphology nor the viability of the cells inside. Conclusion: Microencapsulated engineered CHO cells survive at least 6 weeks after preparation in vitro, and secrete bioactive TIMP-2 freely from the microcapsules.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

[1] Brand, K., Baker, A.H., Perez-Canto, A., Possling, A., Sacharjat, M., Geheeb, M., Arnold, W., 2000. Treatment of colorectal liver metastases by adenoviral transfer of tissue inhibitor of metalloproteinases-2 into the liver tissue. Cancer Res., 60:5723-5730.

[2] Chang, P.L., van Raamsdonk, J.M., Hortelano, G., Barsoum, S.C., MacDonald, N.C., Stockley, T.L., 1999. The in vivo delivery of heterologous proteins by microencapsulated recombinant cells. Trends Biotechnol., 17:78-83.

[3] Cirone, P., Bourgeois, J.M., Chang, P.L., 2003. Antiangiogenic cancer therapy with microencapsulated cells. Hum. Gene Ther., 14:1065-1077.

[4] Hojilla, C.V., Mohammed, F.F., Khokha, R., 2003. Matrix metalloproteinases and their tissue inhibitors direct cell fate during cancer development. Br. J. Cancer, 89:1817-1821.

[5] Imren, S., Kohn, D.B., Shimada, H., Blavier, L., DeClerck, Y.A., 1996. Overexpression of tissue inhibitor of metalloproteinases-2 by retroviral-mediated gene transfer in vivo inhibits tumor growth and invasion. Cancer Res., 56:2891-2895.

[6] Joki, T., Machluf, M., Atala, A., Zhu, J., Seyfried, N.T., Dunn, I.F., Abe, T., Carroll, R.S., Black, P.M., 2001. Continuous release of endostatin from microencapsulated engineered cells for tumor therapy. Nat. Biotechnol., 19:35-39.

[7] Li, H., Lindenmeyer, F., Grenet, C., Opolon, P., Menashi, S., Soria, C., Yeh, P., Perricaudet, M., Lu, H., 2001. AdTIMP-2 inhibits tumor growth, angiogenesis, and metastasis, and prolongs survival in mice. Hum. Gene Ther., 12:515-526.

[8] Lim, F., Sun, A.M., 1980. Microencapsulated islets as bioartificial edocrine pancreas. Science, 210:908-910.

[9] Montgomery, A.M., Mueller, B.M., Reisfeld, R.A., Taylor, S.M., DeClerck, Y.A., 1994. Effect of tissue inhibitor of the matrix metalloproteinases-2 expression on the growth and spontaneous metastasis of a human melanoma cell line. Cancer Res., 54:5467-5473.

[10] Omer, A., Duvivier-Kali, V.F., Trivedi, N., Wilmot, K., Bonner-Weir, S., Weir, G.C., 2003. Survival and maturation of microencapsulated porcine neonatal pancreatic cell clusters transplanted into immunocompetent diabetic mice. Diabetes, 52:69-75.

[11] Orive, G., Hernandez, R.M., Gascon, A.R., Calafiore, R., Chang, T.M., de Vos, P., Hortelano, G., Hunkeler, D., Lacik, I., Shapiro, A.M., Pedraz, J.L., 2003. Cell encapsulation: Promise and progress. Nat. Med., 9:104-107.

[12] Orive, G., Hernandez, R.M., Gascon, A.R., Calafiore, R., Chang, T.M., de Vos, P., Hortelano, G., Hunkeler, D., Lacik, I., Pedraz, J.L., 2004. History, challenges and perspectives of cell microencapsulation. Trends Biotechnol., 22:87-92.

[13] Read, T.A., Sorensen, D.R., Mahesparan, R., Enger, P.O., Timpl, R., Olsen, B.R., Hjelstuen, M.H., Haraldseth, O., Bjerkvig, R., 2001. Local endostatin treatment of gliomas administered by microencapsulated producer cells. Nat. Biotechnol., 19:29-34.

[14] Stetler-Stevenson, W.G., 1999. Matrix metalloproteinases in angiogenesis: A moving target for therapeutic intervention. J. Clin. Invest., 103:1237-1241.

[15] Tai, I.T., Sun, A.M., 1993. Microencapsulation of recombinant cells: A new delivery system for gene therapy. FASEB J., 7:1061-1069.

[16] Uludag, H., de Vos, P., Tresco, P.A., 2000. Technology of mammalian cell encapsulation. Adv. Drug Deliv. Rev., 42:29-64.

[17] Valente, P., Fassina, G., Melchiori, A., Masiello, L., Cilli, M., Vacca, A., Onisto, M., Santi, L., Stetler-Stevenson, W.G., Albini, A., 1998. TIMP-2 over-expression reduces invasion and angiogenesis and protects B16F10 melanoma cells from apoptosis. Int. J. Cancer, 75:246-253.

[18] Vincent, L., Varet, J., Pille, J.Y., Bompais, H., Opolon, P., Maksimenko, A., Malvy, C., Mirshahi, M., Lu, H., Vannier, J.P., Soria, C., Li, H., 2003. Efficacy of dendrimer-mediated angiostatin and TIMP-2 gene delivery on inhibition of tumor growth and angiogenesis: In vitro and in vivo studies. Int. J. Cancer, 105:419-429.

[19] Xu, W., Liu, L., Charles, I.G., 2001. Microencapsulated iNOS-expressing cells cause tumor suppression in mice. FASEB J., 16:213-215.

Open peer comments: Debate/Discuss/Question/Opinion

<1>

Please provide your name, email address and a comment





Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - Journal of Zhejiang University-SCIENCE