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. 2025 Feb 25;9(4):673-686.
doi: 10.1182/bloodadvances.2024014475.

Donor age contributes more to the rheological properties of stored red blood cells than donor sex and biological age distribution

Olga Mykhailova  1 , Mackenzie Brandon-Coatham  1 , Sanaz Hemmatibardehshahi  1   2 , Mahsa Yazdanbakhsh  1   2 , Carly Olafson  1 , Qi-Long Yi  3 , Tamir Kanias  4 , Jason P Acker  1   2
Affiliations

Donor age contributes more to the rheological properties of stored red blood cells than donor sex and biological age distribution

Olga Mykhailova et al. Blood Adv. .

Abstract

The quality of stored red cell concentrates (RCCs) has been linked to the biological age distribution of red blood cell (RBC) subpopulations. Teenage male RCCs contain higher proportions of biologically old RBCs, with poorer quality. This study sought to assess the contribution of donor sex and age on the deformability characteristics of RBC subpopulations in stored RCCs. On days 5, 14, 28, and 42 of hypothermic storage, RCCs from healthy teenage male (n = 15), senior male (n = 15), teenage female (n = 15), and senior female (n = 15) donors were biologically age profiled. The deformability of the resulting young RBCs and old RBCs (O-RBCs) was assessed using ektacytometry. Over storage, donor age was the biggest factor influencing the rheology of RBC subpopulations. Teenage male RCCs had the largest reduction in Ohyper (osmolality in the hypertonic region corresponding to 50% of the maximum RBC elongation [EImax]). The strongest correlations between Ohyper and mean corpuscular hemoglobin content (R2 > 0.5) were witnessed with O-RBCs from senior donors, and to a lesser extent with teenage males. Teen O-RBCs, particularly from males, had higher elongation indices, both under isotonic conditions and in the presence of an increasing osmotic gradient. Teen RBCs, regardless of biological age, were discovered to be more rigid (higher shear stress required to reach half the EImax). Donor variation in the age distribution of RBC subpopulations and its downstream effect on deformability serves as further evidence that factors beyond storage could potentially affect RCC quality and transfusion outcomes.

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Conflict of interest statement

Conflict-of-interest disclosure: The authors declare no competing financial interests.

Figures

None
Graphical abstract
Figure 1.
Figure 1.
Effect of age and sex stratification on the Ohyper of biological age–profiled RCCs. (A) Ohyper measurements for the 3 populations of RBCs (U-RBCs, Y-RBCs, and O-RBCs) examined without any stratification based on donor characteristics. (B-D) Ohyper conveyed as age and sex stratified for U-RBCs (B), Y-RBCs (C), and O-RBCs (D). Multiple comparisons tests were used to show significant differences (∗∗∗∗P < .0001; ∗∗∗P < .001; ∗∗P < .01; ∗P < .05). Median and interquartile ranges (IQR) are indicated as horizontal lines on the violin plots.
Figure 2.
Figure 2.
For U-RBCs, the correlations between Ohyper and MCHC throughout storage and based on donor characteristics. Across hypothermic storage testing points, Ohyper and MCHC correlations for U-RBCs are shown as age and sex stratified into the following donor groups: SF (A), SM (B), TF (C), and TM (D). The corresponding R2 values are provided in Table 2.
Figure 3.
Figure 3.
For O-RBCs, the correlations between Ohyper and MCHC throughout storage and based on donor characteristics. Across hypothermic storage testing points, Ohyper and MCHC correlations for O-RBCs are shown as age and sex stratified into the following donor groups: SF (A), SM (B), TF (C), and TM (D). The corresponding R2 values are provided in Table 3.
Figure 4.
Figure 4.
For Y-RBCs, the correlations between Ohyper and MCHC throughout storage and based on donor characteristics. Across hypothermic storage testing points, Ohyper and MCHC correlations for Y-RBCs are shown as age and sex stratified into the following donor groups: SF (A), SM (B), TF (C), and TM (D). The corresponding R2values are provided in Table 4.
Figure 5.
Figure 5.
Effect of age and sex stratification on multiple elongation measurements taken under an osmotic gradient of biological age–profiled RCCs. (A) Delta elongation measurements for the 3 populations of RBCs (U-RBCs, Y-RBCs, and O-RBCs) without any stratification based on donor characteristics. (B) Delta elongation conveyed as age stratified for O-RBCs only. (C) EIhyper measurements for the 3 populations of RBCs (U-RBCs, Y-RBCs, and O-RBCs) without any stratification based on donor characteristics. (D) EIhyper conveyed as age and sex stratified for O-RBCs only. (E) EImin measurements for the 3 populations of RBCs (U-RBCs, Y-RBCs, and O-RBCs) without any stratification based on donor characteristics. (F) EImin conveyed as age and sex stratified for Y-RBCs only. Multiple comparisons tests were used to show significant differences (∗∗∗∗P < .0001; ∗∗∗P < .001; ∗∗P < .01; ∗P < .05). Median and IQR are indicated as horizontal lines on the violin plots.
Figure 6.
Figure 6.
Effect of age and sex stratification on the EImax of biological age–profiled RCCs. (A) EImax measurements for the 3 populations of RBCs (U-RBCs, Y-RBCs, and O-RBCs) examined without any stratification based on donor characteristics. (B-D) EImax conveyed as age and sex stratified for U-RBCs (B), Y-RBCs (C), and O-RBCs (D). Multiple comparisons tests were used to show significant differences (∗∗∗∗P < .0001; ∗∗∗P < .001; ∗∗P < .01; ∗P < .05). Median and IQR are indicated as horizontal lines on the violin plots.
Figure 7.
Figure 7.
Effect of age and sex stratification on the rigidity (KEI) of biological age–profiled RCCs. (A) KEI measurements for the 3 populations of RBCs (U-RBCs, Y-RBCs, and O-RBCs) examined without any stratification based on donor characteristics. (B-D) KEI conveyed as age and sex stratified for U-RBCs (B), Y-RBCs (C), and O-RBCs (D). Multiple comparisons tests were used to show significant differences (∗∗∗∗P < .0001; ∗∗∗P < .001; ∗∗P < .01; ∗P < .05). Median and IQR are indicated as horizontal lines on the violin plots.
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