Biochemical, biophysical, and genetic changes of porcine trophoblast-derived stem-like cells during differentiation as evaluated using Raman microspectroscopy, Atomic force microscopy, and quantitative polymerase chain reaction.
Porcine trophoblast-derived stem-like cells grown into serum medium start to differentiate and become senescent within 30 days. However, trophoblast-derived cells, cultured in vitro in a defined and non-serum medium, have the regenerative properties, such as indefinite passage and foreign DNA receptivity, similar to stem cells. To evaluate the biochemical, biophysical, and genetic changes of the terminal differentiation of trophoblast derived cells, Raman microspectroscopy, atomic force microscopy, and qPCR were applied. It was found that Raman spectral intensities of characteristic peaks, cell morphology, and Young's modulus can be used to distinguish differentiated and undifferentiated trophoblast cells. In addition, 17 cytoskeleton and extracellular matrix-related genes were significantly impacted by medium type (non-serum versus serum). Our findings suggest that Raman microspectroscopy and atomic force microscopy-both considered as label-free, non-invasive techniques-can be applied to distinguish differentiated trophoblast cells, and cellular biochemical information and biophysical properties can be indicative of cellular differences during cell differentiation. In addition, most of cytoskeleton-related genes exhibit similar pattern to that of Young's modulus during trophoblast cell differentiation, indicating the potential connection between cytoskeleton-related genes and cellular stiffness.
Department of Biological Engineering, Utah State University, Logan, Utah.
Qifei
Q
Li
Department of Animal Science, Dairy, and Veterinary Sciences, Utah State University, Logan, Utah.
Edison
E
Suasnavas
Department of Animal Science, Dairy, and Veterinary Sciences, Utah State University, Logan, Utah.
Sierra
S
Heywood
Department of Biological Engineering, Utah State University, Logan, Utah.
Lifu
L
Xiao
Department of Biological Engineering, Utah State University, Logan, Utah.
Anhong
A
Zhou
Department of Animal Science, Dairy, and Veterinary Sciences, Utah State University, Logan, Utah.
S Clay
SC
Isom
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002462:Cell Membrane / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D018625:Microscopy, Atomic Force / Q000379:methods; D015591:Microspectrophotometry / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D013552:Swine; D014327:Trophoblasts / Q000166:cytology* / Q000378:metabolism
Raman microspectroscopy; atomic force microscopy; biomechanics-related genes; quantitative polymerase chain reaction; trophoblast-derived stem-like cell differentiation