To Determine the Morphological Changes in Red Blood Cells During Storage in Blood Banks

Background: During storage of blood, the red blood cells undergo shape changes which cause fragility and endothelial interaction leading to deterioration the quality of blood in blood banks.Objectives: The aim of this study is to determine the morphological changes in red blood cells during storage in blood banks. Material and Methods: In this experimental study, a total 20 healthy volunteers between 17 to 40 years blood donors-Blood bags were taken, ten from each center i.e. MMCTH blood bank Mardan and KTH blood bank Peshawar. The specimen analysis was done at IBMS (Institute of Basic Medical Sciences) of KMU (Khyber Medical University) Peshawar. The exclusion criteria were People with anemia, hepatitis B &C, HIV and syphilis. The duration of this study was six months. The inform consent was taken from each donor. The total blood 250 ml from vein in cubital fossa from each blood donor was collected in 250ml pediatric blood bag with CPDA-1 solution. Blood bags were put up in the blood bank at +2 to +6 °C and stored till 20 days. Blood specimen of about 5cc were collected in 5cc syringe from each blood bag on 0, 5th,10th ,15th and 20th day for following parameters and thin film red blood cell was prepared for examination by light microscope. Morphological changes in RBCs examined via light microscope as well as grading the RBCs status in the peripheral blood film, the occurrence of distorted RBC simply in random fields; such as +1(scored 1 to 5 altered RBC present in each field), +2 (an average of 6 to 15 altered RBC in each field), +3(16 to 25 altered RBC in each field) and +4(more than 25 altered RBC present in each field). The multi head light microscope NIKON eclipse 50 was used for examination of peripheral blood slide and we took images of randomly selected field. The image J software was used for slide examination.Results: The morphological analysis of red blood cells, count of 200 cells in each blood slide in randomly selected fields are: On day 0 the majority of cells were normally shaped (97.95±1.297 (mean±SD).With increasing storage time, the percentage of morphologically abnormal red cells rose sharply. Mean percentage of abnormal cells on day 5, 10, 15 and 20 was 28.80±10.00, 51.73±12.47, 64.78±14.66 and 68.10±7.92 respectively. This increase in percentage of abnormally shaped cells was significant as determined by one way ANOVA (p =0.001). There was a big difference of percentage of abnormal RBCs on day 0 and in = 5 to= 10 days and in = 15 to = 20 days of blood storage. The mean values of day 0 of abnormal cells was 2.05±1.297 (Mean ± Std. Deviation), abnormal cells in= 5 to= 10 days was 40.26± 16.101 (Mean ± Std. Deviation) and on day = 15 and in = 20 day was 66.44± 11.75. The mean difference from day 0 to day 20 was 63.93±10.45 (Mean ± Std. Deviation).The one way ANOVA was significant, P= 0.001.Conclusion: This study confirms the hematological and morphological changes, when blood stored at 2 °C to 6 °C for up to 21 days. The significant morphological changes were observed on 5th day of blood storage. These findings suggested that approximately a week old stored blood is as good as the fresh blood; however, significant morphological and biochemical changes begin to appear after the first week of storage and these changes aggravate with time. Hence in order to achieve best possible transfusion outcomes, stored blood up to one week can be utilized.

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An Evaluation of Morphological Changes and Deformability of Suspended Red Blood Cells Prepared Using Whole Blood with Different Hemoglobin Levels of Tibetans

Transfusion Medicine and Hemotherapy ◽

10.1159/000513319 ◽

2021 ◽

pp. 1-10

Author(s):

Rui Zhong ◽

Dingding Han ◽

Xiaodong Wu ◽

Hong Wang ◽

Wanjing Li ◽

...

Keyword(s):

Red Blood Cells ◽

Whole Blood ◽

Blood Cells ◽

Morphological Changes ◽

Osmotic Fragility ◽

Spherical Shape ◽

Hypoxic Environment ◽

Wide Range ◽

Group A ◽

Cell Membrane Protein

Background: The hypoxic environment stimulates the human body to increase the levels of hemoglobin (HGB) and hematocrit and the number of red blood cells. Such enhancements have individual differences, leading to a wide range of HGB in Tibetans’ whole blood (WB). Study Design: WB of male Tibetans was divided into 3 groups according to different HGB (i.e., A: >120 but ≤185 g/L, B: >185 but ≤210 g/L, and C: >210 g/L). Suspended red blood cells (SRBC) processed by collected WB and stored in standard conditions were examined aseptically on days 1, 14, 21, and 35 after storage. The routine biochemical indexes, deformability, cell morphology, and membrane proteins were tested. Results: Mean corpuscular volume, adenosine triphosphate, pH, and deformability were not different in group A vs. those in storage (p > 0.05). The increased rate of irreversible morphology of red blood cells was different among the 3 groups, but there was no difference in the percentage of red blood cells with an irreversible morphology after 35 days of storage. Group C performed better in terms of osmotic fragility and showed a lower rigid index than group A. Furthermore, SDS-PAGE revealed similar cross-linking degrees of cell membrane protein but the band 3 protein of group C seemed to experience weaker clustering than that of group A as detected by Western Blot analysis after 35 days of storage. Conclusions: There was no difference in deformability or morphological changes in the 3 groups over the 35 days of storage. High HGB levels of plateau SRBC did not accelerate the RBC change from a biconcave disc into a spherical shape and it did not cause a reduction in deformability during 35 days of preservation in bank conditions.

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Hemoglobin Oxidation in Stored Blood Accelerates Hemolysis and Oxidative Injury to Red Blood Cells

Journal of Laboratory Physicians ◽

10.1055/s-0040-1721156 ◽

2020 ◽

Vol 12 (04) ◽

pp. 244-249

Author(s):

Ibrahim Mustafa ◽

Tameem Ali Qaid Hadwan

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

Morphological Changes ◽

Hemoglobin Concentration ◽

Red Cell Membrane ◽

Rbc Membrane ◽

Hemoglobin Oxidation ◽

Rbc Indices ◽

Methemoglobin Formation ◽

Stored Blood

Abstract Introduction Maintaining blood supply is a challenge in blood banks. Red blood cells (RBCs) stored at 4°C experience issues of biochemical changes due to metabolism of cells, leading to changes collectively referred to as “storage lesions.” Oxidation of the red cell membrane, leading to lysis, contributes to these storage lesions. Methods Blood bags with CPD-SAGM stored at 4°C for 28 days were withdrawn aseptically on days 1, 14, and 28. Hematology analyzer was used to investigate RBC indices. Hemoglobin oxidation was studied through spectrophotometric scan of spectral change. RBC lysis was studied with the help of Drabkin's assay, and morphological changes were observed by light and scan electron microscopy. Results RBCs show progressive changes in morphology echinocytes and spherocytes on day 28. There was 0.85% RBC lysis, an approximately 20% decrease in percentage oxyhemoglobin, and a 14% increase in methemoglobin formation, which shows hemoglobin oxidation on day 28. Conclusions Oxidative damage to RBC, with an increase in storage time was observed in the present study. The observed morphological changes to RBC during the course of increased time shows that there is progressive damage to RBC membrane and a decrease in hemoglobin concentration; percentage RBC lysis is probably due to free hemoglobin and iron.

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Mesenchymal stromal cells can be applied to red blood cells storage as a kind of cellular additive

Bioscience Reports ◽

10.1042/bsr20170676 ◽

2017 ◽

Vol 37 (5) ◽

Cited By ~ 1

Author(s):

Yaozhen Chen ◽

Jing Zhang ◽

Shunli Gu ◽

Dandan Yin ◽

Qunxing An ◽

...

Keyword(s):

Red Blood Cells ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Blood Cells ◽

Storage Duration ◽

Hematopoietic Stem ◽

Diminished Capacity ◽

Blood Banks ◽

Citrate Phosphate

During storage in blood banks, red blood cells (RBCs) undergo the mechanical and metabolic damage, which may lead to the diminished capacity to deliver oxygen. At high altitude regions, the above-mentioned damage may get worse. Thus, more attention should be paid to preserve RBCs when these components need transfer from plain to plateau regions. Recently, we found that mesenchymal stromal cells (MSCs) could rescue from anemia, and MSCs have been demonstrated in hematopoietic stem cells (HSCs) transplantation to reconstitute hematopoiesis in vivo by us. Considering the functions and advantages of MSCs mentioned above, we are trying to find out whether they are helpful to RBCs in storage duration at high altitudes. In the present study, we first found that mice MSCs could be preserved in citrate phosphate dextrose adenine-1 (CPDA-1) at 4 ± 2°C for 14 days, and still maintained great viability, even at plateau region. Thus, we attempted to use MSCs as an available supplement to decrease RBCs lesion during storage. We found that MSCs were helpful to support RBCs to maintain biochemical parameters and kept RBCs function well on relieving anemia in an acute hemolytic murine model. Therefore, our investigation developed a method to get a better storage of RBCs through adding MSCs, which may be applied in RBCs storage as a kind of cellular additive into preservation solution.

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Morphological changes of the red blood cells treated with metal oxide nanoparticles

Toxicology in Vitro ◽

10.1016/j.tiv.2016.08.012 ◽

2016 ◽

Vol 37 ◽

pp. 34-40 ◽

Cited By ~ 7

Author(s):

A.I. Kozelskaya ◽

A.V. Panin ◽

I.A. Khlusov ◽

P.V. Mokrushnikov ◽

B.N. Zaitsev ◽

...

Keyword(s):

Metal Oxide ◽

Red Blood Cells ◽

Blood Cells ◽

Morphological Changes ◽

Metal Oxide Nanoparticles ◽

Oxide Nanoparticles

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Cryopreservation of feline red blood cells in liquid nitrogen using glycerol and hydroxyethyl starch

Journal of Feline Medicine and Surgery ◽

10.1177/1098612x19850932 ◽

2019 ◽

Vol 22 (4) ◽

pp. 366-375

Author(s):

Marcelle Hon ◽

Elizabeth J Thomovsky ◽

Aimee C Brooks ◽

Paula A Johnson

Keyword(s):

Red Blood Cells ◽

Liquid Nitrogen ◽

Hydroxyethyl Starch ◽

Blood Cells ◽

Recovery Rate ◽

Morphological Changes ◽

High Viscosity ◽

Washing Process ◽

Handling Technique ◽

Packed Rbcs

ObjectivesThe objective of this study was to evaluate the techniques and short-term effects of cryopreservation of feline red blood cells (RBCs) in liquid nitrogen using glycerol or hydroxyethyl starch (HES) as a cryoprotectant.MethodsFeline RBCs were manually mixed with either 20% glycerol or 12.5% HES and frozen for 24 h in liquid nitrogen. The samples were thawed and glycerolized samples were manually washed. Success of the freeze/thaw process was determined by recovery rate of RBCs and evaluation of morphological changes using scanning electron microscopy (SEM). A unit of canine packed RBCs was also subjected to the same methodology to evaluate the cryopreservation handling technique.ResultsFeline RBCs preserved with 20% glycerol had a high recovery rate (94.23 ± 1.25%) immediately after thawing. However, the majority of the cells were lost during the washing process, with a final packed cell volume of <1%. A recovery rate was unable to be assessed for samples preserved with HES owing to the high viscosity of the mixture. SEM revealed significant morphological changes after glycerol was added to the feline RBCs. Although these morphological changes were partially reversed after thawing, the majority of the RBCs were lost during the washing process. Minimal morphological changes were noted in the HES sample. Similar results were noted with the canine RBCs.Conclusions and relevanceThe described manual technique for cryopreservation using glycerol was not able to successfully preserve feline or canine RBCs. In the present study, it was difficult to make conclusions about the efficacy of HES. Further studies evaluating HES as a cryoprotectant are warranted.

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