Optimized Enrichment of CD71<sup>+</sup> Reticulocytes From Whole Blood for Single‐Cell RNA Sequencing
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<sup>+</sup> 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.
Authors
- P Okoro (ORCID: https://orcid.org/0009-0005-6589-1101)
- Mohammed Mamman (ORCID: https://orcid.org/0000-0001-5885-1607)
- Halima Bello‐Manga (ORCID: https://orcid.org/0000-0001-9993-4548)
- Abubakar Musa
- Gloria Dada Chechet (ORCID: https://orcid.org/0000-0003-2741-3940)
- Reuben S. Baba
- Idowu A. Aimola (ORCID: https://orcid.org/0000-0001-7553-438X)
- Hanneda A. Fomukong
- Joy U. Ameloko
- Abubakar Sani
Institutions
- Ahmadu Bello University (NG)
- Kaduna State University (NG)
- Hassan Usman Katsina Polytechnic (NG)
- The Federal Polytechnic, Ado-Ekiti (NG)
- Kano State University of Technology (NG)
- Bayero University Kano (NG)
Publication Details
- Journal
- Current Protocols
- Published
- 2025-09-01
- DOI
- https://doi.org/10.1002/cpz1.70203
- Primary Topic
- Single-cell and spatial transcriptomics
- Type
- article
- Field-Weighted Citation Impact
- 0.00