These tools will no longer be maintained as of December 31, 2024. Archived website can be found here. PubMed4Hh GitHub repository can be found here. Contact NLM Customer Service if you have questions.
Pubmed for Handhelds
PUBMED FOR HANDHELDS
Search MEDLINE/PubMed
Title: Biological properties of subpopulations of pluripotent hematopoietic stem cells enriched by elutriation and flow cytometry. Author: Orlic D, Anderson S, Bodine DM. Journal: Blood Cells; 1994; 20(1):107-17; discussion 118-20. PubMed ID: 7527675. Abstract: We have studied several features of pluripotent hematopoietic stem cells (PHSCs) and day-12 spleen colony-forming units (CFU-S) obtained from adult murine bone marrow. Single-cell suspensions of C57BL/6J mouse bone marrow were fractionated by counterflow centrifugal elutriation at flow rates (FR) of 15, 25, 30, and 35 ml/min, and with the rotor off (R/O). The fractions FR25 and FR35 contained approximately equal numbers of PHSC that could repopulate W/Wv mice. These PHSCs were further enriched by subtracting lineage-positive cells using monoclonal antibodies (MAb) and magnetic immunobeads. The resulting lineage-negative cells (Lin-) were then stained with a MAb for the c-kit receptor and sorted by flow cytometry. Both subsets were fractionated into cells expressing high (bright) (c-kitBR), low (dull) c-kitDULL and no (negative, c-kitNEG) c-kit receptor. As few as 100 to 200 c-kitBR cells could repopulate the entire thymus and bone marrow in W/Wv mice. No PHSCs were present in the c-kitDULL and c-kitNEG fractions. We assayed fresh bone marrow and elutriation fractions FR25 and FR35 for gene expression by reverse transcriptase polymerase chain reaction. Using a semiquantitative protocol, we detected mRNA for beta-globin and flk-2, a protein tyrosine kinase receptor, in all samples except the FR25 Lin- c-kitBR subset. We consider the cells in FR25 Lin- c-kitBR to be the most primitive set of hematopoietic stem cells.[Abstract] [Full Text] [Related] [New Search]