Maseko TE

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MiPsociety
COST Action CA15203 MitoEAGLE
Evolution-Age-Gender-Lifestyle-Environment: mitochondrial fitness mapping


 

Maseko TE


MitoPedia topics: EAGLE 

COST: Member


COST WG3: WG3 COST WG4: WG4


Name Maseko Tumisang Edward, Dr.
Institution Faculty of Medicine in Hradec Kralove,

Charles University, CZ

Address Simkova 870, 500 03
City Hradec Kralove
State/Province
Country Czech Republic
Email albacast91@gmail.com
Weblink
O2k-Network Lab CZ Hradec Kralove Cervinkova Z


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Publications

 PublishedReference
Gnaiger 2019 MitoFit Preprint Arch2019Gnaiger E, Aasander Frostner E, Abdul Karim N, Abdel-Rahman EA, Abumrad NA, Acuna-Castroviejo D, Adiele RC, et al (2019) Mitochondrial respiratory states and rates. MitoFit Preprint Arch doi:10.26124/mitofit:190001.v6.

Abstracts

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MitoEAGLE Short-Term Scientific Mission

Work Plan summary
In the month of September 2018, I was working with Dr Jordi Muntane, who was here on STSM. :::: He cultured HepG2 and HepaRG cell lines and treated them with the drug Sorafenib (10 micromolar and 10 nanomolar) as part of his research objectives to show how these concentrations affect mitochondrial functions, Sorafenib 10 nanomolar caused an upregulation of mitochondrial respiration, mitochondrial hyperpolarization and activation of autophagy with reduction of apoptosis in HepG2 cells (24hr) while in 10 micromolar Sorafenib we observed low mitochondrial respiration capacity and mitochondrial depolarization in HepG2 cells ( 24hr). Working in this area has motivated me to further go on to learn and evaluate if mitochondrial dysfunction by Sorafenib is related to alteration of electron transport activity in different complexes. Other objectives that I wish to accomplish in this mission are as follows;
1)Learn the procedure of isolating and culturing primary and liver cancer cells
2)Measure the activities of complex I, II, III, I+II, I+III, and IV
3)Evaluation of coenzyme Q (oxidized and reduced state)
4)Markers of mitophagy using western-blot analysis
5)Strengthen cooperation among groups as part of MiTOEAGLE objectives
6)Learn other experimental procedures that are not available in our Institution
Proposed Contributions to the Scientific Objectives of the Action
The overall objective of the MITOEAGLE network is to improve our knowledge of mitochondrial function in health and disease related to Evolution, Age, Gender, Lifestyle and Environment, as outlined above this mission will improve my knowledge in mitochondrial physiology and related parameters and help contribute to the relevant working groups ( WG3) and further consolidate corporation between the MITOEAGLE network.
Techniques and Equipments to be used
The in vitro experiments will be based on cultured HepG2 and primary hepatocytes. The maintenance of HepG2 cells requires the use of cell culture room fully equipped. Sorafenib will be administered to cell culture and/or in permeabilized cells in suspension at therapeutic dose (10 µM). Mitochondrial activities will be measured by biochemical procedures. The measurement of oxidized and reduced coenzyme Q will be performed using HPLC. The expression of mitophagy or autophagy markers will be evaluated using western-blot analysis.
Working Plan
The experiments can be perfectly done during the scheduled SMTS period (January 8-March 7).
1st phase: January 8-January 25
1) Culture, maintenance and expansion of culture of HepG2 cells.
2) Culturing primary human hepatocytes and collecting samples.
3) Measurement of complex I, II, III, I+II, I+III, and IV activities
2nd phase: January 26-February 27
1) Evaluation of coenzyme Q (oxidized and reduced state)
2) Markers of mitophagy using western-blot analysis
3rd phase: February 28-March 7
1) Evaluation of data and conclusions of the SMTS


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