[4S00m] 01. ASBA - Part 10

10:45 AM - 11:00 AM JST(1:45 AM - 2:00 AM UTC)

[4ASBA-04] Scalable production of reagent red blood cells from a stable erythroid cell line

○Jiahai Shi¹ (1. National University of Singapore (Singapore))

Keywords:

reagent RBC,stable erythroid cell line,blood group antigens,immunohematology

Reagent red blood cells (RBCs) are essential for immunohematology testing, yet donor-derived reagent supply is limited by lot-to-lot variability, finite access to rare phenotypes, and challenges in maintaining consistent antigen performance across sites and time. A scalable, cell-based manufacturing route could improve standardization and enable defined reference materials, but long-term expansion of many erythroid platforms is complicated by karyotypic drift and clonal heterogeneity that undermine product consistency.Here we describe an integrated culture engineering workflow that links a genomically stable erythroid cell line to scalable production of reagent RBCs with consistent antigen performance. We established a suspension-compatible human erythroid line immortalized at a late differentiation stage to preserve erythroid competence while enabling robust, reproducible expansion. Notably, after one year of continuous culture, the line showed no evidence of karyotypic drift or increased chromosome content, supporting long-term manufacturing use. Upon induction, cells followed a reproducible maturation trajectory marked by acquisition of mature erythroid markers, enabling predictable presentation of RBC-relevant surface markers. Building on this reproducibility, we developed a reagent-RBC production process incorporating in-process analytics (maturation state and antigen expression by flow cytometry), and stability evaluation under standard reagent storage conditions. Finally, we outline a route toward programmable reference panels by combining controlled culture with targeted genetic modulation to generate defined antigen-positive and antigen-negative controls, including candidates relevant to clinically important and rare specificities.Together, this work establishes a practical foundation for scalable, quality-controlled reagent RBC manufacturing, enabling more reproducible testing materials and expanding access to defined antigen states for assay development and inter-laboratory harmonization.