Spherocytic shift of red blood cells during storage provides a quantitative whole cell-based marker of the storage lesion.
Storage lesion may explain the rapid clearance of up to 25% of transfused red blood cells (RBCs) in recipients. Several alterations affect stored RBC but a quantitative, whole cell-based predictor of transfusion yield is lacking. Because RBCs with reduced surface area are retained by the spleen, we quantified changes in RBC dimensions during storage.



Using imaging flow cytometry we observed the dimension and morphology of RBCs upon storage, along with that of conventional biochemical and mechanical markers of storage lesion. We then validated these findings using differential interference contrast (DIC) microscopy and quantified the accumulation of microparticles (MPs).



Mean projected surface area of the whole RBC population decreased from 72.4 to 68.4 µm2, a change resulting from the appearance of a well-demarcated subpopulation of RBCs with reduced mean projected surface (58 µm2, 15.2%-19.9% reduction). These "small RBCs" accounted for 4.9 and 23.6% of all RBCs on Days 3 and 42 of storage, respectively. DIC microscopy confirmed that small RBCs had shifted upon storage from discocytes to echinocytes III, spheroechinocytes, and spherocytes. Glycophorin A-positive MPs and small RBCs appeared after similar kinetics.



The reduction in surface area of small RBCs is expected to induce their retention by the spleen. We propose that small RBCs generated by MP-induced membrane loss are preferentially cleared from the circulation shortly after transfusion of long-stored blood. Their operator-independent quantification using imaging flow cytometry may provide a marker of storage lesion potentially predictive of transfusion yield.
Université Sorbonne Paris Cité, Université Paris Descartes, Inserm, INTS, Unité Biologie Intégrée du Globule Rouge.|Université Sorbonne Paris Cité, Université Paris Descartes, Inserm, CNRS, Institut Imagine, Laboratory of Cellular and Molecular Mechanisms of Hematological Disorders and Therapeutic Implications, Laboratoire d'Excellence GR-Ex.
Camille
C
Roussel
Université Sorbonne Paris Cité, Université Paris Descartes, Inserm, CNRS, Institut Imagine, Laboratory of Cellular and Molecular Mechanisms of Hematological Disorders and Therapeutic Implications, Laboratoire d'Excellence GR-Ex.
Michaël
M
Dussiot
Université Sorbonne Paris Cité, Université Paris Diderot, Inserm, INTS, Unité Biologie Intégrée du Globule Rouge.
Mickaël
M
Marin
Université Sorbonne Paris Cité, Université Paris Descartes, Inserm, CNRS, Institut Imagine, Laboratory of Cellular and Molecular Mechanisms of Hematological Disorders and Therapeutic Implications, Laboratoire d'Excellence GR-Ex.
Alexandre
A
Morel
Université Sorbonne Paris Cité, Université Paris Descartes, Inserm, INTS, Unité Biologie Intégrée du Globule Rouge.
Papa Alioune
PA
Ndour
Université Sorbonne Paris Cité, Université Paris Descartes, Inserm, INTS, Unité Biologie Intégrée du Globule Rouge.
Julien
J
Duez
Université Sorbonne Paris Cité, Université Paris Diderot, Inserm, INTS, Unité Biologie Intégrée du Globule Rouge.
Caroline
C
Le Van Kim
Université Sorbonne Paris Cité, Université Paris Descartes, Inserm, CNRS, Institut Imagine, Laboratory of Cellular and Molecular Mechanisms of Hematological Disorders and Therapeutic Implications, Laboratoire d'Excellence GR-Ex.|Department of Adult Hematology, Necker Hospital, Assistance Publique Hôpitaux de Paris, Paris, France.
Olivier
O
Hermine
Université Sorbonne Paris Cité, Université Paris Diderot, Inserm, INTS, Unité Biologie Intégrée du Globule Rouge.
Yves
Y
Colin
Université Sorbonne Paris Cité, Université Paris Descartes, Inserm, INTS, Unité Biologie Intégrée du Globule Rouge.
Pierre A
PA
Buffet
Université Sorbonne Paris Cité, Université Paris Diderot, Inserm, INTS, Unité Biologie Intégrée du Globule Rouge.|Université Sorbonne Paris Cité, Université Paris Descartes, Inserm, CNRS, Institut Imagine, Laboratory of Cellular and Molecular Mechanisms of Hematological Disorders and Therapeutic Implications, Laboratoire d'Excellence GR-Ex.
Pascal
P
Amireault
D015415:Biomarkers / Q000097:blood; D001793:Blood Preservation; D055252:Cell-Derived Microparticles; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007700:Kinetics; D008297:Male; D013102:Spherocytes / Q000166:cytology* / Q000378:metabolism; D013997:Time Factors
D015415:Biomarkers