Production and stability of cultured red blood cells depends on the concentration of cholesterol in culture medium.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
06 Jul 2024
06 Jul 2024
Historique:
received:
22
04
2024
accepted:
01
07
2024
medline:
7
7
2024
pubmed:
7
7
2024
entrez:
6
7
2024
Statut:
epublish
Résumé
The production of cultured red blood cells (cRBC) for transfusion purposes requires large scale cultures and downstream processes to purify enucleated cRBC. The membrane composition, and cholesterol content in particular, are important during proliferation of (pro)erythroblasts and for cRBC quality. Therefore, we tested the requirement for cholesterol in the culture medium during expansion and differentiation of erythroid cultures with respect to proliferation, enucleation and purification by filtration. The low cholesterol level (22 µg/dl) in serum free medium was sufficient to expand (pro)erythroblast cultures. Addition of 2.0 or 5.0 mg/dL of free cholesterol at the start of differentiation induction inhibited enucleation compared to the default condition containing 3.3 mg/dl total cholesterol derived from the addition of Omniplasma to serum free medium. Addition of 5.0 mg/dl cholesterol at day 5 of differentiation did not affect the enucleation process but significantly increased recovery of enucleated cRBC following filtration over leukodepletion filters. The addition of cholesterol at day 5 increased the osmotic resistance of cRBC. In conclusion, cholesterol supplementation after the onset of enucleation improved the robustness of cRBC and increased the yield of enucleated cRBC in the purification process.
Identifiants
pubmed: 38971841
doi: 10.1038/s41598-024-66440-z
pii: 10.1038/s41598-024-66440-z
doi:
Substances chimiques
Cholesterol
97C5T2UQ7J
Culture Media
0
Culture Media, Serum-Free
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
15592Subventions
Organisme : ZonMw
ID : TAS project 116003004
Pays : Netherlands
Organisme : ZonMw
ID : TAS project 116003004
Pays : Netherlands
Organisme : ZonMw
ID : TAS project 116003004
Pays : Netherlands
Organisme : Sanquin Blood Supply
ID : PPOC17-28
Informations de copyright
© 2024. The Author(s).
Références
Knowles, S. Blood transfusion: Challenges and limitations. Transfus. Altern. Transfus. Med. 9, 2–9 (2007).
doi: 10.1111/j.1778-428X.2007.00062.x
Landsteiner, K. Zur Kenntnis der antifermentativen, lytischen und agglutinierenden Wirkungen des Blutserums und der Lymphe. Zentralblatt fur Bakteriol. Parasitenkd. und Infekt. 27, 357–362 (1900).
Landsteiner, K. & Wiener, A. S. Studies on an agglutinogen (Rh) in human blood reacting with anti-rhesus sera and with human isoantibodies. J. Exp. Med. 74, 309–320 (1941).
doi: 10.1084/jem.74.4.309
Gassner, C. et al. International society of blood transfusion working party on red cell immunogenetics and blood group terminology report of basel and three virtual business meetings: Update on blood group systems. Vox Sang. 117, 1332–1344 (2022).
doi: 10.1111/vox.13361
Pirenne, F., Floch, A. & Diop, S. Alloimmunisation against red blood cells in sickle cell disease: Transfusion challenges in high-income and low-income countries. Lancet Haematol. 10, e468–e476 (2023).
doi: 10.1016/S2352-3026(23)00066-2
Giarratana, M. C. et al. Proof of principle for transfusion of in vitro-generated red blood cells. Blood 118, 5071–5079 (2011).
doi: 10.1182/blood-2011-06-362038
Hawksworth, J. et al. Enhancement of red blood cell transfusion compatibility using CRISPR-mediated erythroblast gene editing. EMBO Mol. Med. 10, e8454 (2018).
doi: 10.15252/emmm.201708454
Masiello, F. et al. Mononuclear cells from a rare blood donor, after freezing under good manufacturing practice conditions, generate red blood cells that recapitulate the rare blood phenotype. Transfusion 54, 1059–1070 (2014).
doi: 10.1111/trf.12391
Migliaccio, A. R., Grazzini, G. & Hillyer, C. D. Ex vivo generated red cells as transfusion products. Stem Cells Int. 2012, 615412 (2012).
doi: 10.1155/2012/615412
Kupzig, S., Parsons, S. F., Curnow, E., Anstee, D. J. & Blair, A. Superior survival of ex vivo cultured human reticulocytes following transfusion into mice. Haematologica 102, 476–483 (2017).
doi: 10.3324/haematol.2016.154443
van den Akker, E., Satchwell, T. J., Pellegrin, S., Daniels, G. & Toye, A. M. The majority of the in vitro erythroid expansion potential resides in CD34- cells, outweighing the contribution of CD34+ cells and significantly increasing the erythroblast yield from peripheral blood samples. Haematologica 95, 1594–1598 (2010).
doi: 10.3324/haematol.2009.019828
Leberbauer, C. et al. Different steroids co-regulate long-term expansion versus terminal differentiation in primary human erythroid progenitors. Blood 105, 85–94 (2005).
doi: 10.1182/blood-2004-03-1002
Heshusius, S. et al. Large-scale in vitro production of red blood cells from human peripheral blood mononuclear cells. Blood Adv. 3, 3337–3350 (2019).
doi: 10.1182/bloodadvances.2019000689
Migliaccio, G. et al. Humanized culture medium for clinical expansion of human erythroblasts. Cell Transplant. 19, 453–469 (2010).
doi: 10.3727/096368909X485049
Gallego-Murillo, J. S. et al. Expansion and differentiation of ex vivo cultured erythroblasts in scalable stirred bioreactors. Biotechnol. Bioeng. https://doi.org/10.1002/bit.28193 (2022).
doi: 10.1002/bit.28193
Glen, K. E., Cheeseman, E. A., Stacey, A. J. & Thomas, R. J. A mechanistic model of erythroblast growth inhibition providing a framework for optimisation of cell therapy manufacturing. Biochem. Eng. J. 133, 28–38 (2018).
doi: 10.1016/j.bej.2018.01.033
Higgins, V. L. Leukocyte-reduced blood components: patient benefits and practical applications. Oncol. Nurs. Forum 23, 659–667 (1996).
Mohandas, N. & Gallagher, P. G. Red cell membrane: Past, present, and future. Blood 112, 3939–3948 (2008).
doi: 10.1182/blood-2008-07-161166
van den Akker, E., Satchwell, T. J., Williamson, R. C. & Toye, A. M. Band 3 multiprotein complexes in the red cell membrane; of mice and men. Blood Cells. Mol. Dis. 45, 1–8 (2010).
doi: 10.1016/j.bcmd.2010.02.019
Brewer, G. J. Inherited erythrocyte metabolic and membrane disorders. Med. Clin. N. Am. 64, 579–596 (1980).
doi: 10.1016/S0025-7125(16)31582-6
An, X. & Mohandas, N. Disorders of red cell membrane. Br. J. Haematol. 141, 367–375 (2008).
doi: 10.1111/j.1365-2141.2008.07091.x
Pollet, H., Conrard, L., Cloos, A. S. & Tyteca, D. Plasma membrane lipid domains as platforms for vesicle biogenesis and shedding?. Biomolecules 8, 94 (2018).
doi: 10.3390/biom8030094
Bernecker, C. et al. Cholesterol deficiency causes impaired osmotic stability of cultured red blood cells. Front. Physiol. 10, 1–14 (2019).
doi: 10.3389/fphys.2019.01529
van den Broek, I., Sobhani, K. & Van Eyk, J. E. Advances in quantifying apolipoproteins using LC-MS/MS technology: Implications for the clinic. Expert Rev. Proteom. 14, 869–880 (2017).
doi: 10.1080/14789450.2017.1374859
Subczynski, W. K., Pasenkiewicz-Gierula, M., Widomska, J., Mainali, L. & Raguz, M. High cholesterol/low cholesterol: Effects in biological membranes: A review. Cell Biochem. Biophys. 75, 369–385 (2017).
doi: 10.1007/s12013-017-0792-7
Samkari, A. et al. A novel missense mutation in MVK associated with MK deficiency and dyserythropoietic anemia. Pediatrics 125, 964–968 (2010).
doi: 10.1542/peds.2009-1774
Holm, T. M. et al. Failure of red blood cell maturation in mice with defects in the high-density lipoprotein receptor SR-BI. Blood 99, 1817–1824 (2002).
doi: 10.1182/blood.V99.5.1817.h8001817_1817_1824
Oram, J. O. Molecular basis of cholesterol homeostasis: Lessons from Tangier disease and ABCA1. Trends Mol. Med. 8, 168–173 (2002).
doi: 10.1016/S1471-4914(02)02289-X
Atac, B., Brahaj, D., Frishman, W. H. & Lerner, R. Anemia and hypocholesterolemia. Heart Dis. 5, 65–71 (2003).
doi: 10.1097/01.HDX.0000050420.79522.09
Huang, N. J. et al. Enhanced phosphocholine metabolism is essential for terminal erythropoiesis. Blood 131, 2955–2966 (2018).
doi: 10.1182/blood-2018-03-838516
Migliaccio, G. & Migliaccio, A. R. Cloning of human erythroid progenitors (BFU-E) in the absence of fetal bovine serum. Br. J. Haematol. 67, 129–133 (2008).
Baek, E. J., Kim, H., Kim, J., Kim, N. J. & Kim, H. O. Stroma-free mass production of clinical-grade red blood cells (RBCs) by using poloxamer 188 as an RBC survival enhancer. Transfusion 49, 2285–2295 (2009).
doi: 10.1111/j.1537-2995.2009.02303.x
Griffiths, R. E. et al. Maturing reticulocytes internalize plasma membrane in glycophorin A–containing vesicles that fuse with autophagosomes before exocytosis. Blood 119, 6296–6306 (2012).
doi: 10.1182/blood-2011-09-376475
Miharada, K., Hiroyama, T., Sudo, K., Nagasawa, T. & Nakamura, Y. Efficient enucleation of erythroblasts differentiated in vitro from hematopoietic stem and progenitor cells. Nat. Biotechnol. 24, 1255–1256 (2006).
doi: 10.1038/nbt1245
Neildez-Nguyen, T. M. A. et al. Human erythroid cells produced ex vivo at large scale differentiate into red blood cells in vivo. Nat. Biotechnol. 20, 467–472 (2002).
doi: 10.1038/nbt0502-467
Gold, J. C. & Phillips, M. C. Effects of membrane lipid composition on the kinetics of cholesterol exchange between lipoproteins and different species of red blood cells. Biochim. Biophys. Acta 1027, 85–92 (1990).
doi: 10.1016/0005-2736(90)90052-P
Schick, B. P. & Schick, P. K. Cholesterol exchange in platelets, erythrocytes and megakaryocytes. Biochim. Biophys. Acta 833, 281–290 (1985).
doi: 10.1016/0005-2760(85)90200-0
Gottlieb, M. H. Rates of cholesterol exchange between human erythrocytes and plasma lipoproteins. Biochim. Biophys. Acta 600, 530–541 (1980).
doi: 10.1016/0005-2736(80)90454-X
Ohkawa, R. et al. Cholesterol transport between red blood cells and lipoproteins contributes to cholesterol metabolism in blood. J. Lipid Res. 61, 1577–1588 (2020).
doi: 10.1194/jlr.RA120000635
Solanko, L. M. et al. Membrane orientation and lateral diffusion of BODIPY-cholesterol as a function of probe structure. Biophys. J. 105, 2082–2092 (2013).
doi: 10.1016/j.bpj.2013.09.031
Hölttä-Vuori, M., Sezgin, E., Eggeling, C. & Ikonen, E. Use of BODIPY-cholesterol (TF-Chol) for visualizing lysosomal cholesterol accumulation. Traffic 17, 1054–1057 (2016).
doi: 10.1111/tra.12414
Zhang, K., Cai, R., Chen, D. & Mao, L. Determination of hemoglobin based on its enzymatic activity for the oxidation of o-phenylenediamine with hydrogen peroxide. Anal. Chim. Acta 413, 109–113 (2000).
doi: 10.1016/S0003-2670(00)00752-2
Dobbe, J. G. G., Streekstra, G. J., Hardeman, M. R., Ince, C. & Grimbergen, C. A. Measurement of the distribution of red blood cell deformability using an automated rheoscope. Cytometry 50, 313–325 (2002).
doi: 10.1002/cyto.10171
van Zwieten, R. et al. Partial pyruvate kinase deficiency aggravates the phenotypic expression of band 3 deficiency in a family with hereditary spherocytosis. Am. J. Hematol. 90, E35–E39 (2015).
Paul, R., Zhou, Y., Nikfar, M., Razizadeh, M. & Liu, Y. Quantitative absorption imaging of red blood cells to determine physical and mechanical properties. RSC Adv. 10, 38923–38936 (2020).
doi: 10.1039/D0RA05421F
Chen, K. et al. Resolving the distinct stages in erythroid differentiation based on dynamic changes in membrane protein expression during erythropoiesis. Proc. Natl. Acad. Sci. 106, 17413–17418 (2009).
doi: 10.1073/pnas.0909296106
Iacono, G. et al. Differentiating erythroblasts adapt to turbulent flow by accelerating maturation and activating cholesterol biosynthesis. BioRxiv, https://doi.org/10.1101/2023.12.08.570773 (2023)
Patel, R., Williams-Dautovich, J. & Cummins, C. L. Minireview: New molecular mediators of glucocorticoid receptor activity in metabolic tissues. Mol. Endocrinol. 28, 999–1011 (2014).
doi: 10.1210/me.2014-1062
Zingariello, M. et al. Dexamethasone predisposes human erythroblasts toward impaired lipid metabolism and renders their ex vivo expansion highly dependent on plasma lipoproteins. Front. Physiol. 10, 281 (2019).
doi: 10.3389/fphys.2019.00281
Wang, E. et al. An optimized human erythroblast differentiation system reveals cholesterol-dependency of robust production of cultured red blood cells ex vivo. Adv. Sci. https://doi.org/10.1002/advs.202303471 (2024).
doi: 10.1002/advs.202303471
Buchwald, H. et al. Plasma cholesterol: An influencing factor in red blood cell oxygen release and cellular oxygen availability. J. Am. Coll. Surg. 191, 490–497 (2000).
doi: 10.1016/S1072-7515(00)00704-3
Ridone, P. et al. Disruption of membrane cholesterol organization impairs the activity of PIEZO1 channel clusters. J. Gen. Physiol. 152, e201912515 (2020).
doi: 10.1085/jgp.201912515
Luneva, O. G. et al. Ion transport, membrane fluidity and haemoglobin conformation in erythrocyte from patients with cardiovascular diseases: Role of augmented plasma cholesterol. Pathophysiol. Off. J. Int. Soc. Pathophysiol. 14, 41–46 (2007).
Leonard, C. et al. Contribution of plasma membrane lipid domains to red blood cell (re)shaping. Sci. Rep. 7, 4264 (2017).
doi: 10.1038/s41598-017-04388-z
Hung, K. T., Berisha, S. Z., Ritchey, B. M., Santore, J. & Smith, J. D. Red blood cells play a role in reverse cholesterol transport. Arterioscler. Thromb. Vasc. Biol. 32, 1460–1465 (2012).
doi: 10.1161/ATVBAHA.112.248971
Hayashi, R. et al. Serum cholesterol level in normal people association of serum cholesterol level with age and relative body weight. Jpn. J. Med. 26, 153–157 (1987).
doi: 10.2169/internalmedicine1962.26.153
Loi, M. M. et al. A comparison of different methods of red blood cell leukoreduction and additive solutions on the accumulation of neutrophil-priming activity during storage. Transfusion 58, 2003–2012 (2018).
doi: 10.1111/trf.14788
Singh, S. & Kumar, A. Leukocyte depletion for safe blood transfusion. Biotechnol. J. 4, 1140–1151 (2009).
doi: 10.1002/biot.200800182
Trampler, F., Sonderhoff, S. A., Pui, P. W., Kilburn, D. G. & Piret, J. M. Acoustic cell filter for high density perfusion culture of hybridoma cells. Biotechnology 12, 281–284 (1994).
doi: 10.1038/nbt0394-281
Zeming, K. K. et al. Microfluidic label-free bioprocessing of human reticulocytes from erythroid culture. Lab Chip 20, 3445–3460 (2020).
doi: 10.1039/C9LC01128E
Pellegrin, S., Severn, C. E. & Toye, A. M. Towards manufactured red blood cells for the treatment of inherited anemia. Haematologica 106, 2304–2311 (2021).
doi: 10.3324/haematol.2020.268847
Karst, D. J., Serra, E., Villiger, T. K., Soos, M. & Morbidelli, M. Characterization and comparison of ATF and TFF in stirred bioreactors for continuous mammalian cell culture processes. Biochem. Eng. J. 110, 17–26 (2016).
doi: 10.1016/j.bej.2016.02.003