Biochemical analysis and quantification of hematopoietic stem cells by infrared spectroscopy.
Identification of hematopoietic stem cells (HSCs) in different stages of maturation is one of the major issues in stem cell research and bone marrow (BM) transplantation. Each stage of maturation of HSCs is characterized by a series of distinct glycoproteins present on the cell plasma membrane surface, named a cluster of differentiation (CD). Currently, complicated and expensive procedures based on CD expression are needed for identification and isolation of HSCs. This method is under dispute, since the correct markers' composition is not strictly clear, thus there is need for a better method for stem cell characterization. In the present study, Fourier transform infrared (FTIR) spectroscopy is employed as a novel optical method for identification and characterization of HSCs based on their entire biochemical features. FTIR spectral analysis of isolated mice HSCs reveals several spectral markers related to lipids, nucleic acids, and carbohydrates, which distinguish HSCs from BM cells. The unique "open" conformation of HSC DNA as identified by FTIR is exploited for HSCs quantification in the BM. The proposed method of FTIR spectroscopy for HSC identification and quantification can contribute to stem cell research and BM transplantation.
Zelig
Udi
U
Ben-Gurion University of the Negev, Department of Biomedical Engineering, Beer-Sheva 84105 Israel.
Dror
Ziv
Z
Iskovich
Svetlana
S
Zwielly
Amir
A
Ben-Harush
Miri
M
Nathan
Ilana
I
Mordechai
Shaul
S
Kapelushnik
Joseph
J
D000818:Animals; D015415:Biomarkers / Q000737:chemistry; D001854:Bone Marrow Cells / Q000737:chemistry; D004247:DNA / Q000737:chemistry; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000737:chemistry*; D016018:Least-Squares Analysis; D051379:Mice; D008810:Mice, Inbred C57BL; D008853:Microscopy / Q000379:methods*; D009690:Nucleic Acid Conformation; D025341:Principal Component Analysis; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*
D015415:Biomarkers; D004247:DNA