comparison of three methods for mitochondria isolation from the human liver cell line (hepg2)

نویسندگان

pedram azimzadeh reproductive biotechnology research center, avicenna research institute, acecr, tehran, iran

hamid asadzadeh aghdaei basic and molecular epidemiology of gastrointestinal disorders, gastroenterology and liver diseases research institute, shahid beheshti university of medical sciences, tehran, iran

peyman tarban basic and molecular epidemiology of gastrointestinal disorders, gastroenterology and liver diseases research institute, shahid beheshti university of medical sciences, tehran, iran

mohammad mahdi akhondi reproductive biotechnology research center, avicenna research institute, acecr, tehran, iran

چکیده

aim : the aim of this study was to evaluate and compare three available methods for mitochondrial isolation from a human cell line to predict the best method for each probable application.   background : organelle isolation is gaining importance in experimental laboratory settings. mitochondrial dysfunction may affect tumorgenesis process. there are some evidences that transplantation of healthy, intact and active mitochondria into cells containing defective mitochondria may reduce the proliferation. therefore, isolated mitochondria could be considered as an effective tool for assessment and management of mitochondrial related disorders. patients and methods : mitochondrial isolation from the human liver cell line ( hepg2 ) was performed using two commercially available kits, including qproteome (qiagen) and mitoiso2 (sigma-aldrich), as well as a manual method. integrity of inner membrane of mitochondria was assessed by jc-1 staining. activity of isolated mitochondria was evaluated by dcfh-da staining, and total yield of isolated mitochondria determined by micro-lowry method. finally, relative quantification using real-time pcr was conducted to compare the mtdna copy number of mitochondria isolated by three different methods. results : compared to other methods, manual kit resulted in higher yields of total amount of mitochondrial protein and mtdna copy numbers. isolated mitochondria by qproteome kit, showed a higher activity. finally, the integrity of inner-membrane of isolated mitochondria was significantly higher in qproteome when compared with the other two methods. conclusion : due to differences in quality, quantity and activity of isolated mitochondria using three techniques discussed here, the method in which best-suited to each research project should be selected according to the distinct features of isolated mitochondria.

برای دانلود باید عضویت طلایی داشته باشید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Comparison of three methods for mitochondria isolation from the human liver cell line (HepG2).

AIM The aim of this study was to evaluate and compare three available methods for mitochondrial isolation from a human cell line to predict the best method for each probable application. BACKGROUND Organelle isolation is gaining importance in experimental laboratory settings. Mitochondrial dysfunction may affect tumorgenesis process. There are some evidences that transplantation of healthy, i...

متن کامل

Toxicity Effect of Silver Nanoparticles on Mice Liver Primary Cell Culture and HepG2 Cell Line

Nano-silver (AgNP) has biological properties which are significant for consumer products, food technology, textiles and medical applications (e.g. wound care products, implantable medical devices, in diagnosis, drug delivery, and imaging). For their antibacterial activity, silver nanoparticles are largely used in various commercially available products. Thus, the use of nano-silver is becoming ...

متن کامل

Toxicity Effect of Silver Nanoparticles on Mice Liver Primary Cell Culture and HepG2 Cell Line

Nano-silver (AgNP) has biological properties which are significant for consumer products, food technology, textiles and medical applications (e.g. wound care products, implantable medical devices, in diagnosis, drug delivery, and imaging). For their antibacterial activity, silver nanoparticles are largely used in various commercially available products. Thus, the use of nano-silver is becoming ...

متن کامل

on the comparison of keyword and semantic-context methods of learning new vocabulary meaning

the rationale behind the present study is that particular learning strategies produce more effective results when applied together. the present study tried to investigate the efficiency of the semantic-context strategy alone with a technique called, keyword method. to clarify the point, the current study seeked to find answer to the following question: are the keyword and semantic-context metho...

15 صفحه اول

Antioxidant Effects of Pinus Eldarica Extracts Against Cisplatin-Induced Cytotoxicity in Human Hepatoma (Hepg2) Cell Line

Background:  Cisplatin has cytotoxic effects through free radical generation. Reports indicate that various members of Pinaceae family may have antioxidant properties. In this study we investigated the cytoprotective effects of needle volatile oil and bark extract of Pinus eldarica in hepatoma G2 (HepG2) cell line.  Methods: Using the maceration technique, we obtained the ethanolic extract of ...

متن کامل

Cytotoxicity of Subtoxic AgNP in Human Hepatoma Cell Line (HepG2) after Long-Term Exposure

Background: We aimed at evaluating the toxicity effects of low (subtoxic) concentrations of silver nanoparticles (AgNP, 5-10 nm) in human hepatoblastoma (HepG2) cell line after and during a period of about one month. Methods: XTT and MTT assays were used to draw a dose-response curve IC50 (half maximal inhibitory concentration) value of the AgNP on HepG2 cells was calculated to be 2.75-3.0 mg/l...

متن کامل

منابع من

با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید


عنوان ژورنال:
gastroenterology and hepatology from bed to bench

جلد ۹، شماره ۲، صفحات ۲۰۱۶-۹

کلمات کلیدی

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023