Laser-assisted cell removing (LACR) technology contributes to the purification process of the undifferentiated cell fraction during pluripotent stem cell culture

Biochem Biophys Res Commun. 2018 Sep 18;503(4):3114-3120. doi: 10.1016/j.bbrc.2018.08.101. Epub 2018 Aug 22.

Abstract

Purification of undifferentiated cells by removing differentiated parts is an essential step in pluripotent stem cell culture. This process has been traditionally performed manually using a fine glass capillary or plastic tip under a microscope, or by culturing in a selective medium supplemented with anti-differentiation inhibitors. However, there are several inevitable problems associated with these methods, such as contamination or biological side-effects. Here, we developed a laser-assisted cell removing (LACR) technology that enables precise, fast, and contact-less cell removal. Using LACR combined with computational image recognition/identification-discriminating technology, we achieved automatic cell purification (A-LACR). Practicability of A-LACR was evaluated by two demonstrations: selective removal of trophoblast stem (TS) cells from human iPS and TS cell co-cultures, and purification of undifferentiated iPS cells by targeting differentiated cells that spontaneously developed. Our results suggested that LACR technology is a novel approach for stem cell processing in regenerative medicine.

Keywords: Automated cell culture; NIR laser; iPS cells.

MeSH terms

  • Animals
  • Cell Culture Techniques / methods*
  • Cell Death / radiation effects
  • Cell Differentiation
  • Cell Line
  • Coculture Techniques / methods
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / radiation effects
  • Infrared Rays / adverse effects
  • Lasers / adverse effects
  • Mice
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / radiation effects
  • Regenerative Medicine
  • Trophoblasts / cytology*
  • Trophoblasts / radiation effects