Microscopy To Measure The Potency Of Stem Cells

Researchers in Japan have developed a modified version of differential interference contrast microscopy which helps identify pluripotent stem cells with the greatest medical potential.

AsianScientist (Apr. 10, 2019) – In a study published in Scientific Reports, a research group in Japan has developed a microscope that can identify pluripotent stem cells (PSCs) with the highest likelihood of differentiating into desired tissues.

PSCs can transform into any type of cell in the human body. This unique property gives them enormous potential for regenerative medicine, as stem cells might eventually be used to replace tissues that are lost as a result of degenerative disorders or from injuries. Not all stem cells are created equal, though, and some have greater differentiation potential than others.

Traditionally, a rigorous process is needed to weed out potentially problematic cells from the medically beneficial ones. This often involves staining cells or labeling them with special fluorescent markers—invasive procedures that can damage or destroy the stem cells.

In the present study, researchers led by Professor Koji Hisatake of the University of Tsukuba, Japan, have sought to identify PSCs ideal for use in regenerative therapy.

“Cells that have low pluripotency are not especially effective after being transplanted in a patient. Even worse, in some cases there’s a risk that they can form tumors. Our goal was to improve on earlier microscopy techniques so that we can do a better job at isolating stem cells with high pluripotency, which are more likely to be safe and effective for therapeutic use,” said Professor Koji Hisatake of the University of Tsukuba, Japan.

The researchers thus developed a modified form of differential interference contrast (DIC) microscopy, which they called phase distribution (PD) imaging. The technique allowed the team to visualize cell structures based on differences in light scattering. Unlike DIC, the crystal prisms that polarize light waves are movable in the PD microscope, which means that two separate images of the cell, at right angles from one another, are captured at once, then merged into a single image using a computer.

The researchers noted that the final output image had a much greater level of detail than traditional DIC microscopes. Using the PD microscope, the researchers were able to distinguish energy-generating mitochondria within cells and found that highly potent PSCs had fewer and smaller mitochondria. By quantifying mitochondria in each PSC, they were able to reliably isolate PSCs with the greatest medical potential.

“PD imaging allows us to non-invasively identify stem cells that have been fully reprogrammed and likely have exceptional pluripotency,” said Dr. Ken Nishimura of the University of Tsukuba, the lead author of the study. “The system is less costly and time-consuming than other screening methods, so we’re hopeful that this will eventually become an important tool in the screening arsenal, especially during the critical early stages of stem cell selection.”



The article can be found at: Nishimura et al. (2019) Live-cell Imaging of Subcellular Structures for Quantitative Evaluation of Pluripotent Stem Cells.

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Source: University of Tsukuba; Photo: Shutterstock.
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