Multiparameter analysis of transplantable hemopoietic stem cells. I. The separation and enrichment of stem cells homing to marrow and spleen on the basis of rhodamine-123 fluorescence
A multiparameter cell separative procedure is described that enables normal transplantable hemopoietic stem cells that preferentially home to the marrow of lethally irradiated mice to be enriched and separated from the majority of spleen colony-forming cells that are assayed 13 days after transplantation (CFU-S13). First, bone marrow cells are centrifuged in a discontinuous bovine serum albumin gradient. Low-density cells are harvested and labeled with the supravital cationic fluorochrome rhodamine 123 (Rh123). Labeled cells are analyzed using a fluorescence-activated cell sorter, and cells are sorted on the basis of relative Rh123 fluorescence within a predetermined forward versus 90 degrees red light scatter window that has been optimized for the recovery and enrichment of cells with marrow repopulating ability (MRA). Cells with MRA were characterized by relatively low Rh123 fluorescence and could be separated from a fraction that fluoresced more intensely and contained the majority of CFU-S13 but low MRA. Cells with platelet repopulating ability cofractionate with MRA whereas cells with erythroid repopulating ability remain associated with CFU-S13.
- Research Organization:
- Cancer Institute, Melbourne, Victoria, Australia
- OSTI ID:
- 6254612
- Journal Information:
- Exp. Hematol.; (United States), Vol. 10
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
BONE MARROW CELLS
FLUORESCENCE
IMMUNOSUPPRESSION
RADIOINDUCTION
CELL FLOW SYSTEMS
CENTRIFUGATION
CYTOLOGICAL TECHNIQUES
LETHAL IRRADIATION
MICE
RHODAMINES
SPLEEN
AMINES
ANIMAL CELLS
ANIMALS
BODY
CARBOXYLIC ACIDS
CONNECTIVE TISSUE CELLS
DYES
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
IRRADIATION
LUMINESCENCE
MAMMALS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
ORGANS
REAGENTS
RODENTS
SEPARATION PROCESSES
SOMATIC CELLS
VERTEBRATES
560152* - Radiation Effects on Animals- Animals
550300 - Cytology