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

Somatic cells can be reprogrammed into induced pluripotent stem cells (iPSCs) by defined factors. Here, we describe a protocol for imaging mitochondrial permeability transition pore (mPTP) opening in reprogramming of somatic cells using a confocal microscope. We also describe a method to sort high and low mPTP opening somatic cells by calcein fluorescence and reprogram these sorted cells to iPSCs. These protocols are also suitable for imaging mPTP opening and uncovering the mechanisms of mPTP function in other cell fate conversions. For complete details on the use and execution of this protocol, please refer to Ying et al. (2018). [Display omitted] •A protocol for imaging the opening of the mitochondrial permeability transition pore (mPTP)•Analysis of mPTP opening during reprogramming•Sorting of the high and low mPTP opening somatic cells Somatic cells can be reprogrammed into induced pluripotent stem cells (iPSCs) by defined factors. Here, we describe a protocol for imaging mitochondrial permeability transition pore (mPTP) opening in reprogramming of somatic cells using a confocal microscope. We also describe a method to sort high and low mPTP opening somatic cells by calcein fluorescence and reprogram these sorted cells to iPSCs. These protocols are also suitable for imaging mPTP opening and uncovering the mechanisms of mPTP function in other cell fate conversions.