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Multiple red cell volumes (RCV) can be independently determined in sheep in vivo using red cells (RBCs) labeled at 4 biotin densities
Author(s) -
Mock Donald M,
Matthews Nell I,
Strauss Ronald G,
Burmeister Leon F,
Schmidt Robert,
Widness John A
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.795.12
Subject(s) - biotin , biotinylation , in vivo , biology , chromatography , chemistry , microbiology and biotechnology , biochemistry , genetics
Here we report our further development and validation of a method for independent, simultaneous measurement of RCV in sheep using flow cytometric enumeration of multiple populations of RBCs labeled at distinct biotin densities. Method The density of biotin on the surface of the RBCs was incrementally increased to produce 4 discrete populations of biotinylated RBCs (BioRBCs). BioRBCs and RBCs labeled with 14 C‐cyanate were infused into the sheep, and blood samples were drawn at 10 min. Results At biotinylation reagent concentrations < 96 µg/mL RBC, the biotin method reproducibly determined RCV; mean error = 4.2% versus the reference 14 C‐cyanate method (n=2 sheep). RCV values agreed among the densities; mean CV= 2% for 4 sheep. For reagent concentrations > 96 µg/mL RBC, BioRBCs were rapidly removed from circulation; this rapid removal caused an overestimate in RCV. Conclusions BioRBCs at densities low enough to survive in the circulation can be used for simultaneous in vivo measurements of RCV in sheep. Speculation In humans, BioRBCs can be used to repeatedly measure RCV in the same individual. This method will be useful in the investigation of the physiology and pathophysiology of the pregnant woman, fetus, and infant because only small sample volumes are required and because the individual is not exposed to radiation. NIH P01 HL046925 and Thrasher Research Foundation 02825‐3.