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ISSN 2410-7751 (Print)
ISSN 2410-776X (Online)

 no6 2014

"Biotechnologia Acta" v. 7, no 6, 2014
https://doi.org/10.15407/biotech7.06.051
Р. 51-56, Bibliography 12, English
Universal Decimal classification: 612.419.014.1:612.119

PHENOTYPING AND SORTING OF MURINE BONE MARROW HAEMATOPOIETIC STEM CELLS  USING  FLOW CYTOMETRY

Kyryk V. M.

State Institute of Genetic and Regenerative Medicine of the National Academy of Medical Sciences of Ukraine, Kyiv

To develop a protocol of multiparametric phenotyping and sorting of LSK-subpopulations of hematopoietic stem cells and to determine their relative numbers in the bone marrow of mice was the goal of this research.  The  modified  protocol  of  multiparametric  phenotyping  of  murine  hematopoietic  stem  cells enable to determine the content of Lin–Sca-1+ c-kit+, Lin–Sca-1+c-kit+flt3+CD150–, Lin–Sca-1+c-kit+flt3+CD150+ and Lin–Sca-1+c-kit+flt3–CD150– subpopulations in bone marrow of FVB mice. It was shown that the dominant population among LSK-cells represents the phenotype Lin–Sca-1+c-kit+flt3–CD150– (57.2 ± 6.8%),  which  characterizes  the  short-term  hematopoietic  stem  cells  responsible  for  myelopoiesis.

Also the protocol of sorting of murine bone marrow LSK-cells was proposed and its effectiveness for subsequent transplantation in experiments was demonstrated. At repeated phenotyping of sorted cells the purity of Lin–Sca-1+c-kit+ cell fraction was 96.6 ± 1.8% with viability up to 89.6 ± 4.6%.

Key words: haematopoietic stem cells, multiparametric flow cytometry, cell sorting.

© Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, 2014