Quantification of Mouse Lymphohematopoietic Stem CellsUsing IR-Spectroscopy

Abstract

Stem cells and in particular, Hematopoietic stem cells (HSCs) therapy is valued for its potential to restore damaged or degenerating tissues. Transplantation of HSCs is already a routine procedure for treatment of blood and immune system disorders. Till now complicated and expensive procedures were needed for identification and isolation of HSC. In the present research we propose a simple and reagent-free procedure for identifying and quantifying lymphohematopoietic stem cells (LHSC) using Fourier transform infrared (FTIR) spectroscopy, which is able to monitor content and conformation of all bio-molecules in cells simultaneously. B6D2F1 mice, 6 to 12 weeks of age were used for this study. Bone marrow (BM) from male B6D2F1 mice was drawn. Putative LHSC were isolated from the BM by common methods according to cell size, markers and high levels of aldehyde dehydrogenase. After isolation of LHSC, various dilutions of LHSC among BM cells were performed and measured under FTIR-microscope. Using spectral and statistical analysis, we identified several spectral markers which characterize and distinct LHSC from pure BM. These markers refer to nucleic acids which increase in LHSC due to their extensive proliferation and differentiation. Furthermore, we achieved high sensitivity quantification of LHSC from BM. (about one LHSC per thousand BM cells) Thus, we conclude that the proposed method of FTIR spectroscopy identification and quantification of LHSC may significantly contribute to stem cells research and hearten the use of stem cells in medicine.