Preparation and in vitro studies of microencapsulated cells releasing human tissue inhibitor of metalloproteinase-2.
نویسندگان
چکیده
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 - approximately 600 microm. 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.
منابع مشابه
Preparation and evaluation of 67Ga-DOTA-Bombesin (7-14) as a tumor scintigraphic agent
Introduction: Bombesin is a 14-aminoacid peptide isolated from frog skin. The mammalian counterparts of the frog peptide are neuromedin B (NMB) and gastrin-releasing peptide (GRP). Bombesin (BBN) is a peptide showing high affinity for the gastrin releasing peptide receptor (GRPr). Prostate, small cell lung cancer, breast, gastric, and colon cancers are known to over...
متن کاملExpression of Matrix Metalloproteinase-2/9 and Tissue Inhibitor of Metalloproteinase-1/2 as Predictive Factors in Oropharyngeal Squamous Cell Carcinoma
Introduction: Metalloproteinases and their tissue inhibitors play an important role in the metastases formation. A multistage process of carcinogenesis requires the involvement of numerous enzymes and compounds that facilitate the expansion of tumor cells. The formation of metastases depends on both metalloproteinases and tissue inhibitors activation leading to the acti...
متن کاملA Concept Suggestion on the Effect of Cigarette Smoking in Inflammatory Destruction of Gingiva
Dear Editor, Cigarette smoking has destructive effect on periodontal tissue. The rates of loss of periodontal attachment and recession of gingival are higher in smokers than non-smokers (1-2). Previous studies on the inflammatory immune responses in smokers’ periodontitis have mainly focused on the role of neutrophils. Tumor necrosis factor–α, prostaglandin E2 and matrix metalloproteinase...
متن کاملIsolation and in vitro cultivation of human skin Keratinocytes and preparation of epidermal sheet
متن کامل
Isolation and culture of human endometrial derived cells as an in vitro model for future implantation studies
Introduction: Monthly regeneration of endometrium after cyclical mensturation confirmed the ability of specific population of the cells that presence in the basalis layer and undergone consecutive hormonal changes that could prepared the endometrial layer for probable implantation. These cells, known as, stem cell. The aim of this study was the isolation and culture of human endometrial derive...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Journal of Zhejiang University. Science. B
دوره 6 9 شماره
صفحات -
تاریخ انتشار 2005