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Optimized Enrichment of CD71+ Reticulocytes From Whole Blood for Single-Cell RNA Sequencing
Hanneda A Fomukong1,2,3, Idowu A Aimola1,2, Abubakar Sani1,2,4
1Department of Biochemistry, Ahmadu Bello University, Zaria, Nigeria.
Abstract:
Reticulocytes are a group of immature red blood cells (RBCs) produced in the bone marrow and released into the bloodstream where they mature into RBCs within 1 to 2 days. They play a crucial role in assessing bone health and many hematological diseases. Isolating viable reticulocytes for use in single-cell transcriptomics has been a challenge due to their similarity to mature RBCs. Their low concentration and continuous maturation to RBCs with time also poses a great challenge. In this article, we optimized a method for enrichment of CD71+ reticulocytes from whole blood, with a high yield and viability for downstream studies. © 2025 Wiley Periodicals LLC. Basic Protocol 1: Blood tissue preparation and reticulocyte enrichment Basic Protocol 2: Visualization and determination of cell viability Alternate Protocol: Counting cells using a hemocytometer.
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