Protocols for analysis of mitochondrial permeability transition pore opening in mouse somatic cell reprogramming

Area of Science:

• Cell Biology
• Stem Cell Research
• Mitochondrial Biology

Background:

• Somatic cells can be reprogrammed into induced pluripotent stem cells (iPSCs) using specific factors.
• Mitochondrial function, particularly the opening of the mitochondrial permeability transition pore (mPTP), is implicated in cellular processes like reprogramming.

Purpose of the Study:

• To establish a protocol for visualizing mPTP opening in somatic cells undergoing reprogramming.
• To develop a method for sorting cells based on their mPTP opening status to enhance iPSC generation.

Main Methods:

• Confocal microscopy was employed to image mPTP opening during somatic cell reprogramming.
• Calcein fluorescence was utilized to sort somatic cells with high and low mPTP opening activity.

Main Results:

• The study successfully demonstrated a method to image mPTP opening in real-time during cell reprogramming.
• Sorting cells based on mPTP opening levels allowed for the selection of cells with potentially higher reprogramming capacity.

Conclusions:

• The described protocols enable the study of mPTP dynamics in cellular reprogramming and other cell fate conversions.
• Understanding mPTP function can provide insights into optimizing reprogramming efficiency and cellular plasticity.