Study Overview
We have introduced the XFe96 analyzer (Agilent Technologies), which can check how cells make and use energy in real time.
1. Checking mitochondrial activity
The XF Stress Kit measures important parts of mitochondrial activity, including:
・Maximum energy-producing ability
・Basic energy production
・ATP production
・Energy loss during the process
2. Checking glycolysis activity
The XF Glycolytic Rate Kit measures how actively cells produce energy by breaking down glucose.
It can evaluate:
・Extra glycolysis ability when cells need more energy
・Normal glycolysis activity
3. Checking ATP production
The XF Real-Time ATP Rate Kit measures:
・Total ATP production in cells
・ATP made by mitochondria
・ATP made through glycolysis
This allows us to understand how cells produce energy.
4. Checking which energy sources cells use
The XF Mito Fuel Flex Kit analyzes how cells use the three main energy sources:
・Glucose (sugar)
・Fatty acids (fat)
・Glutamine (an amino acid)
It can measure:
・Which energy source cells mainly use under normal conditions
・ How easily cells can switch to another energy source
・How strongly cells depend on each energy source
Simple explanation of terms
1. What is ATP?
ATP is a substance that stores and provides energy for living cells.
It is used for many important cell activities.
2. What is mitochondrial respiration?
Mitochondria are parts of cells that make large amounts of ATP using oxygen.
During this process, oxygen is consumed to produce energy.
3. What is glycolysis?
Glycolysis is a process in which cells break down glucose to make ATP.
This process happens outside mitochondria, in the cell fluid.
During glycolysis, cells release lactate and hydrogen ions, which make the area around the cells more acidic.
Basic principles
・Mitochondria use oxygen when making energy. (OCR)
・During glycolysis, the area around cells becomes more acidic. (ECAR)
Equipment used
XFe96 (Agilent Technologies)
Features of the extracellular flux analyzer
■ Real-time measurement of metabolism in living cells
■ Simultaneous evaluation of mitochondrial respiration (OXPHOS) and glycolysis
■ Functional metabolic evaluation using chemical compounds
■ High-throughput
Typical analysis examples
・Cancer research
Evaluating metabolic changes characteristic of cancer cells, etc.
・Immunology Research
Evaluating pro-inflammatory and anti-inflammatory macrophages
Evaluating T cell activation, etc.
・Obesity Research
Evaluating the differentiation and function of brown and beige adipocytes
Evaluating the energy substrates used by cells, etc.
・Mitochondrial Function Assessment
Evaluation of mitochondrial function by measuring basal and maximal mitochondrial respiration, etc.
References
●Selective Cancer Therapy Effects of Vitamin C
Vitamin C selectively kills KRAS and BRAF mutant colorectal cancer cells by targeting GAPDH
Yun et al., Science. 2015
●CAR-T Cancer Treatment
Targeting REGNASE-1 programs long-lived effector T cells for cancer therapy
Wei et al., Nature. 2019
●iPSC Survival, Differentiation, and Substrate Dependence (Selectivity)
Glutamine Oxidation is indespensable for Survival of Human Pluripotent Stem Cells
Tohyama et al., Cell Metabolism. 2016
*Studies will be mainly conducted by those with doctoral degrees.
