Enhanced Production of Red Blood Cells in Suspension by Electrostatic Interactions with Culture Plates

Abstract

Enucleation of erythroblasts, a critical step in the generation of red blood cells (RBCs), occurs at a low rate without cocultured stromal cells. Previously, the surface properties of the cell culture plate were not considered in the enucleation process, because the cells exist in suspension. Here, we show that a significantly higher rate of enucleation of erythroblasts occurred on the positively charged plates than on the negatively charged surfaces or the both negatively and positively charged plates. Also, the negatively and positively charged plate group showed a significantly higher enucleation than did the hydrophobic plates. Therefore, the plates fully coated with amine groups generated 1.88 times more enucleated RBCs than did the hydrophobic plates. This study suggests an important insight into the effect of surface characteristics of cell culture plates on suspension cell culture. Further, this simple and inexpensive procedure could contribute to a more efficient RBC production system, eliminating the need for robust and expensive coculture procedures.

Characterization and comparison of N‐, O‐, and N+O‐functionalized polymer surfaces for efficient (HUVEC) endothelial cell colonization

“…The hybrid (N‐ and O‐) plasma treated Primaria™ was found to display even slightly (but statistically significant, p < 0.05) better performance than TCP, suggesting that the combination of O‐ and N‐containing moieties might constitute the new “gold standard” for cell culture dishes. This would agree with earlier‐published observations by several groups …”

Poly-L-Lysine Increases theEx VivoExpansion and Erythroid Differentiation of Human Hematopoietic Stem Cells, as Well as Erythroid Enucleation Efficacy

“…Baek et al 12 similarly reported that an amine-rich versus an uncharged substrate augmented the number of enucleated erythroid cells, and hypothesized that electrostatic interactions between the cell membrane and the positively charged substrate were responsible. In addition, the investigators suggested an enucleation mechanism by which the direction of enucleation was upward, but no directly supportive evidence was provided.…”

“…10,11 Furthermore, surfaces incorporating primary versus tertiary amines more effectively induced enucleation during the erythroid differentiation of HSCs. 12 However, highly cytotoxic 3-aminopropyl trimethoxylsilane (APES) was utilized during the amine group incorporation. In addition, the amine group density was not optimized for HSC expansion and enucleation during erythroid differentiation, and the underlying mechanism for the interaction between the HSCs and the investigated substrates remains unclear.…”

Large‐scale production of red blood cells from stem cells: What are the technical challenges ahead?

“…Recently, Baek et al [87] proposed an ingenious method to enhance enucleation rates by positively charging the tissue culture plates. These publications emphasize the importance of identifying the best material for erythroid cultures and show that targeted modifications of the material can improve the results.…”