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. 2021 Oct 1;106(10):2779-2782.
doi: 10.3324/haematol.2021.278895.

Sex-dependent membranopathy in stored human red blood cells

Ewa Szczesny-Malysiak  1 , Tasnim Mohaissen  2 , Katarzyna Bulat  1 , Magdalena Kaczmarska  1 , Aleksandra Wajda  3 , Katarzyna M Marzec  4
Affiliations

Sex-dependent membranopathy in stored human red blood cells

Ewa Szczesny-Malysiak et al. Haematologica. .
No abstract available

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Figures

Figure 1.
Figure 1.
Storage time-dependent changes in biochemical, morphological and functional red blood cell parameters. Biochemical parameters: (A) cholesterol, (B) triglycerides, (C) lactate dehydrogenase (LDH), (D) free iron of human packed red blood cells (pRBC), (E) glucose, (F) lactate; chosen protein expression: (G) CD47 and (H) phosphatidylserine (PS); (I) pRBC deformability; the blood count including: (J) mean corpuscular volume (MCV), (K) hematocrit (HCT), (L) mean corpuscular hemoglobin concentration (MCHC). The blood was withdrawn from men (n=12) and women (n=12). Results (A to F) were obtained from pRBC supernatant with ABX Pentra 400 (Horiba Medical, Japan) are given as mean ± standard deviation (SD), data distribution is presented as box plots (mean value, mean ± SD, min-max whiskers). Results (G to L) presented as box plots (median, Q1, Q3, interquartile range, min-max whiskers). Q1, Q3 indicate 25th and 75th percentiles, respectively. Results (G and H) were obtained using a mixture of mouse anti-human antibodies carrying fluorescent markers: CD45 – APC-Cy7 (1:200, BD cat. no. 348815), CD71 – APC (1:200, BD cat. no. 551374), CD47 – PE (1:200, BD cat. no. 556046) and annexin V – FITC (1:200, BD cat. no. 556547). Cellular analysis was performed with LSRII flow cytometer (BD Biosciences). RBC deformability was studied at 37°C as elongation index at shear rates of 20 Pa with the use of ektacytometer RheoScanAnD 300 (RheoMeditech, Seoul, Korea) while parameters (J and L) with hematology analyzer Abc Vet (Horiba Medical, France). Data (A to L) normality was assessed using Shapiro-Wilk test. The significance of the differences between men and women in each week was evaluated by One-Way ANOVA with Tukey’s test. If data distribution was not normal, non-parametric Kruskal-Wallis test with the Dunn’s post hoc test if appropriate; *P<0.05, **P<0.01, ***P<0.001 ****P<0.0001 indicates significant difference between men and women in each week.
Figure 2.
Figure 2.
Time sequence of the appearance of red blood cell (RBC)-derived microparticles on the surface of human packed RBC isolated from (A) men and (B) women. Examples of atomic force microscopy (AFM) images with the use of WITec confocal CRM alpha 300 in non-contact mode (AC) (WITec, Ulm, Germany) and dry Zeiss objective (ECEPIPLAN 20x/0.4). AFM images of 256x256 lines and 512x512 lines were collected from areas of 25x25 mm2, 8x8 mm2 and 1.5x1.5 mm2, which were performed at room temperature on dried smears of (A) female RBC and (B) male RBC fixed with 1% glutaraldehyde (10 minutes). (C) Sex-specific, time-dependent changes of the RBC-derived microparticle (RMP) sizes observed on the surface of human packed RBC. Data distribution is presented as box plots (median, Q1, Q3, interquartile range, min-max whiskers). Q1, Q3 indicate 25th and 75th percentiles, respectively. *Weeks 7 and 8 are additional measurements exceeding expiration date. Statistical significance of the obtained results (n>35) was tested with Kruskal-Wallis ANOVA non-parametric test followed by Tukey’s post hoc. NS: not significant; *P<0.05; **P<0.01, ***P<0.001, ****P<0.0001).
Figure 3.
Figure 3.
Schematic summary of the sex-specific, time-dependent changes in packed red blood cells derived from men (n=12) and women (n=12). Packed red blood cells (pRBC) were cold-stored for 8 weeks (weeks 7 and 8 being additional time points exceeding expiration date). Physiological parameters were assessed with use of biochemical analysis, morphological analysis, flow cytometry, ektacytometry and atomic force microscopy. fRBC: female RBC; mRBC: male RBC; LDH: lactate dehydrogenase; RMP: red blood cell-derived mircoparticle.
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