Abstract
Erythropoiesis is a process during which multipotential hematopoietic stem cells proliferate, differentiate and eventually form mature erythrocytes. Interestingly, unlike most cell types, an important feature of erythropoiesis is that following each mitosis the daughter cells are morphologically and functionally different from the parent cell from which they are derived, demonstrating the need to study erythropoiesis in a stage-specific manner. This has been impossible until recently due to lack of methods for isolating erythroid cells at each distinct developmental stage. This review summarizes recent advances in the development of methods for isolating both murine and human erythroid cells and their applications. These methods provide powerful means for studying normal and impaired erythropoiesis associated with hematological disorders.
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Palis J. Ontogeny of erythropoiesis. Curr Opin Hematol 2008, 15: 155–161
Finch CA, Sturgeon P. Erythrokinetics in cooley’s anemia. Blood 1957, 12: 64–73
Pootrakul P, Sirankapracha P, Sankote J, Kachintorn U, Maungsub W, Sriphen K, Thakernpol K, Atisuk K, Fucharoen S, Chantraluksri U, Shalev O, Hoffbrand AV. Clinical trial of deferiprone iron chelation therapy in beta-thalassaemia/haemoglobin E patients in thailand. Br J Haematol 2003, 122: 305–310
Kean LS, Brown LE, Nichols JW, Mohandas N, Archer DR, Hsu LL. Comparison of mechanisms of anemia in mice with sickle cell disease and beta-thalassemia: peripheral destruction, ineffective erythropoiesis, and phospholipid scramblase-mediated phosphatidylserine exposure. Exp Hematol 2002, 30: 394–402
Ginzburg Y, Rivella S. Beta-thalassemia: a model for elucidating the dynamic regulation of ineffective erythropoiesis and iron metabolism. Blood 2011, 118: 4321–4330
Wickramasinghe SN, Wood WG. Advances in the understanding of the congenital dyserythropoietic anaemias. Br J Haematol 2005, 131: 431–446
Heimpel H, Schwarz K, Ebnother M, Goede JS, Heydrich D, Kamp T, Plaumann L, Rath B, Roessler J, Schildknecht O, Schmid M, Wuillemin W, Einsiedler B, Leichtle R, Tamary H, Kohne E. Congenital dyserythropoietic anemia type I (CDA I): molecular genetics, clinical appearance, and prognosis based on long-term observation. Blood 2006, 107: 334–340
Schwarz K, Iolascon A, Verissimo F, Trede NS, Horsley W, Chen W, Paw BH, Hopfner KP, Holzmann K, Russo R, Esposito MR, Spano D, De Falco L, Heinrich K, Joggerst B, Rojewski MT, Perrotta S, Denecke J, Pannicke U, Delaunay J, Pepperkok R, Heimpel H. Mutations affecting the secretory COPII coat component SEC23B cause congenital dyserythropoietic anemia type II. Nat Genet 2009, 41: 936–940
Diamond LK, Wang WC, Alter BP. Congenital hypoplastic anemia. Adv Pediatr 1976, 22: 349–378
Lipton JM, Ellis SR. Diamond-Blackfan anemia: diagnosis, treatment, and molecular pathogenesis. Hematol Oncol Clin North Am 2009, 23: 261–282
Chang KH, Tam M, Stevenson MM. Inappropriately low reticulocytosis in severe malarial anemia correlates with suppression in the development of late erythroid precursors. Blood 2004, 103: 3727–3735
Casals-Pascual C, Kai O, Cheung JO, Williams S, Lowe B, Nyanoti M, Williams TN, Maitland K, Molyneux M, Newton CR, Peshu N, Watt SM, Roberts DJ. Suppression of erythropoiesis in malarial anemia is associated with hemozoin in vitro and in vivo. Blood 2006, 108: 2569–2577
Haldar K, Mohandas N. Malaria, erythrocytic infection, and anemia. Hematology, 2009, 87–93
Nimer SD. Myelodysplastic syndromes. Blood 2008, 111: 4841–4851
List A, Dewald G, Bennett J, Giagounidis A, Raza A, Feldman E, Powell B, Greenberg P, Thomas D, Stone R, Reeder C, Wride K, Patin J, Schmidt M, Zeldis J, Knight R, Myelodysplastic Syndrome-003 Study I. Lenalidomide in the myelodysplastic syndrome with chromosome 5q deletion. N Engl J Med 2006, 355: 1456–1465
Ebert BL, Galili N, Tamayo P, Bosco J, Mak R, Pretz J, Tanguturi S, Ladd-Acosta C, Stone R, Golub TR, Raza A. An erythroid differentiation signature predicts response to lenalidomide in myelodysplastic syndrome. PLoS Med, 2008, 5: e35
Lipton JM, Kudisch M, Gross R, Nathan DG. Defective erythroid progenitor differentiation system in congenital hypoplastic (Diamond-Blackfan) anemia. Blood 1986, 67: 962–968
Bagnara GP, Zauli G, Vitale L, Rosito P, Vecchi V, Paolucci G, Avanzi GC, Ramenghi U, Timeus F, Gabutti V. In vitro growth and regulation of bone marrow enriched CD34+ hematopoietic progenitors in diamond-blackfan anemia. Blood 1991, 78: 2203–2210
Dgany O, Avidan N, Delaunay J, Krasnov T, Shalmon L, Shalev H, Eidelitz-Markus T, Kapelushnik J, Cattan D, Pariente A, Tulliez M, Cretien A, Schischmanoff PO, Iolascon A, Fibach E, Koren A, Rossler J, Le Merrer M, Yaniv I, Zaizov R, Ben-Asher E, Olender T, Lancet D, Beckmann JS, Tamary H. Congenital dyserythropoietic anemia type I is caused by mutations in codanin-1. Am J Hum Genet 2002, 71: 1467–1474
Arnaud L, Saison C, Helias V, Lucien N, Steschenko D, Giarratana MC, Prehu C, Foliguet B, Montout L, de Brevern AG, Francina A, Ripoche P, Fenneteau O, Da Costa L, Peyrard T, Coghlan G, Illum N, Birgens H, Tamary H, Iolascon A, Delaunay J, Tchernia G, Cartron JP. A dominant mutation in the gene encoding the erythroid transcription factor KLF1 causes a congenital dyserythropoietic anemia. Am J Hum Genet 2010, 87: 721–727
Satchwell TJ, Pellegrin S, Bianchi P, Hawley BR, Gampel A, Mordue KE, Budnik A, Fermo E, Barcellini W, Stephens DJ, van den Akker E, Toye AM. Characteristic phenotypes associated with congenital dyserythropoietic anemia (type II) manifest at different stages of erythropoiesis. Haematologica 2013, 98: 1788–1796
Liljeholm M, Irvine AF, Vikberg AL, Norberg A, Month S, Sandstrom H, Wahlin A, Mishima M, Golovleva I. Congenital dyserythropoietic anemia type III (CDA III) is caused by a mutation in kinesin family member, KIF23. Blood 2013, 121: 4791–4799
Yuan J, Angelucci E, Lucarelli G, Aljurf M, Snyder LM, Kiefer CR, Ma L, Schrier SL. Accelerated programmed cell death (apoptosis) in erythroid precursors of patients with severe beta-thalassemia (cooley’s anemia). Blood 1993, 82: 374–377
Pootrakul P, Sirankapracha P, Hemsorach S, Moungsub W, Kumbunlue R, Piangitjagum A, Wasi P, Ma L, Schrier SL. A correlation of erythrokinetics, ineffective erythropoiesis, and erythroid precursor apoptosis in thai patients with thalassemia. Blood 2000, 96: 2606–2612
Mathias LA, Fisher TC, Zeng L, Meiselman HJ, Weinberg KI, Hiti AL, Malik P. Ineffective erythropoiesis in beta-thalassemia major is due to apoptosis at the polychromatophilic normoblast stage. Exp Hematol 2000, 28: 1343–1353
Pecci A, Travaglino E, Klersy C, Invernizzi R. Apoptosis in relation to CD34 antigen expression in normal and myelodysplastic bone marrow. Acta Haematol 2003, 109: 29–34
Parker JE, Mufti GJ, Rasool F, Mijovic A, Devereux S, Pagliuca A. The role of apoptosis, proliferation, and the Bcl-2-related proteins in the myelodysplastic syndromes and acute myeloid leukemia secondary to MDS. Blood 2000, 96: 3932–3938
Chen K, Liu J, Heck S, Chasis JA, An X, Mohandas N. Resolving the distinct stages in erythroid differentiation based on dynamic changes in membrane protein expression during erythropoiesis. Proc Natl Acad Sci USA 2009, 106: 17413–17418
Liu J, Zhang J, Ginzburg Y, Li H, Xue F, De Franceschi L, Chasis JA, Mohandas N, An X. Quantitative analysis of murine terminal erythroid differentiation in vivo: novel method to study normal and disordered erythropoiesis. Blood, 2013, 121: e43–e49
Hu J, Liu J, Xue F, Halverson G, Reid M, Guo A, Chen L, Raza A, Galili N, Jaffray J, Lane J, Chasis JA, Taylor N, Mohandas N, An X. Isolation and functional characterization of human erythroblasts at distinct stages: implications for understanding of normal and disordered erythropoiesis in vivo. Blood 2013, 121: 3246–3253
Li J, Hale J, Bhagia P, Xue F, Chen L, Jaffray J, Yan H, Lane J, Gallagher PG, Mohandas N, Liu J, An X. Isolation and transcriptome analyses of human erythroid progenitors: BFU-E and CFU-E. Blood 2014, 124: 3636–3645
Stephenson JR, Axelrad AA, McLeod DL, Shreeve MM. Induction of colonies of hemoglobin-synthesizing cells by erythropoietin in vitro. Proc Natl Acad Sci USA 1971, 68: 1542–1546
McLeod DL, Shreeve MM, Axelrad AA. Improved plasma culture system for production of erythrocytic colonies in vitro: quantitative assay method for CFU-E. Blood 1974, 44: 517–534
Iscove NN, Sieber F, Winterhalter KH. Erythroid colony formation in cultures of mouse and human bone marrow: analysis of the requirement for erythropoietin by gel filtration and affinity chromatography on agarose-concanavalin A. J Cell Physiol 1974, 83: 309–320
Moriyama Y, Fisher JW. Effects of testosterone and erythropoietin on erythroid colony formation in human bone marrow cultures. Blood 1975, 45: 665–670
Gregory CJ, Eaves AC. Human marrow cells capable of erythropoietic differentiation in vitro: definition of three erythroid colony responses. Blood 1977, 49: 855–864
Gregory CJ, Eaves AC. Three stages of erythropoietic progenitor cell differentiation distinguished by a number of physical and biologic properties. Blood 1978, 51: 527–537
Flygare J, Rayon Estrada V, Shin C, Gupta S, Lodish HF. HIF1alpha synergizes with glucocorticoids to promote BFU-E progenitor self-renewal. Blood 2011, 117: 3435–3444
Liu J, Mohandas N, An X. Membrane assembly during erythropoiesis. Curr Opin Hematol 2011, 18: 133–138
Suragani RN, Cadena SM, Cawley SM, Sako D, Mitchell D, Li R, Davies MV, Alexander MJ, Devine M, Loveday KS, Underwood KW, Grinberg AV, Quisel JD, Chopra R, Pearsall RS, Seehra J, Kumar R. Transforming growth factor-beta superfamily ligand trap ACE-536 corrects anemia by promoting late-stage erythropoiesis. Nat Med 2014, 20: 408–414
Pimentel H, Parra M, Gee S, Ghanem D, An X, Li J, Mohandas N, Pachter L, Conboy JG. A dynamic alternative splicing program regulates gene expression during terminal erythropoiesis. Nucleic Acids Res 2014, 42: 4031–4042
Blanc L, Ciciotte SL, Gwynn B, Hildick-Smith GJ, Pierce EL, Soltis KA, Cooney JD, Paw BH, Peters LL. Critical function for the Ras-GTPase activating protein RASA3 in vertebrate erythropoiesis and megakaryopoiesis. Proc Natl Acad Sci USA 2012, 109: 12099–12104
Watanabe S, De Zan T, Ishizaki T, Yasuda S, Kamijo H, Yamada D, Aoki T, Kiyonari H, Kaneko H, Shimizu R, Yamamoto M, Goshima G, Narumiya S. Loss of a Rho-regulated actin nucleator, mDia2, impairs cytokinesis during mouse fetal erythropoiesis. Cell Rep 2013, 5: 926–932
Stowell SR, Henry KL, Smith NH, Hudson KE, Halverson GR, Park JC, Bennett AM, Girard-Pierce KR, Arthur CM, Bunting ST, Zimring JC, Hendrickson JE. Alloantibodies to a paternally derived RBC KEL antigen lead to hemolytic disease of the fetus/newborn in a murine model. Blood 2013, 122: 1494–1504
Palis J. Primitive and definitive erythropoiesis in mammals. Front Physiol 2014, 5: 3
Heath DS, Axelrad AA, McLeod DL, Shreeve MM. Separation of the erythropoietin-responsive progenitors BFU-E and CFU-E in mouse bone marrow by unit gravity sedimentation. Blood 1976, 47: 777–792
Terszowski G, Waskow C, Conradt P, Lenze D, Koenigsmann J, Carstanjen D, Horak I, Rodewald HR. Prospective isolation and global gene expression analysis of the erythrocyte colony-forming unit (CFU-E). Blood 2005, 105: 1937–1945
Stumpf M, Waskow C, Krotschel M, van Essen D, Rodriguez P, Zhang X, Guyot B, Roeder RG, Borggrefe T. The mediator complex functions as a coactivator for GATA-1 in erythropoiesis via subunit MED1/TRAP220. Proc Natl Acad Sci USA 2006, 103: 18504–18509
Koury MJ. Abnormal erythropoiesis and the pathophysiology of chronic anemia. Blood Rev 2014, 28: 49–66
Arlet JB, Ribeil JA, Guillem F, Negre O, Hazoume A, Marcion G, Beuzard Y, Dussiot M, Moura IC, Demarest S, de Beauchene IC, Belaid-Choucair Z, Sevin M, Maciel TT, Auclair C, Leboulch P, Chretien S, Tchertanov L, Baudin-Creuza V, Seigneuric R, Fontenay M, Garrido C, Hermine O, Courtois G. Hsp70 sequestration by free alpha-globin promotes ineffective erythropoiesis in beta- thalassaemia. Nature 2014, 514: 242–246
Ludwig LS, Gazda HT, Eng JC, Eichhorn SW, Thiru P, Ghazvinian R, George TI, Gotlib JR, Beggs AH, Sieff CA, Lodish HF, Lander ES, Sankaran VG. Altered translation of GATA1 in Diamond- Blackfan anemia. Nat Med 2014, 20: 748–753
Bibikova E, Youn MY, Danilova N, Ono-Uruga Y, Konto-Ghiorghi Y, Ochoa R, Narla A, Glader B, Lin S, Sakamoto KM. Tnf-mediated inflammation represses GATA1 and activates p38 map kinase in RPS19-deficient hematopoietic progenitors. Blood 2014, 124: 3791–3798
Dai CH, Krantz SB, Zsebo KM. Human burst-forming units-erythroid need direct interaction with stem cell factor for further development. Blood 1991, 78: 2493–2497
Muta K, Krantz SB, Bondurant MC, Wickrema A. Distinct roles of erythropoietin, insulin-like growth factor I, and stem cell factor in the development of erythroid progenitor cells. J Clin Invest 1994, 94: 34–43
Keller MA, Addya S, Vadigepalli R, Banini B, Delgrosso K, Huang H, Surrey S. Transcriptional regulatory network analysis of developing human erythroid progenitors reveals patterns of coregulation and potential transcriptional regulators. Physiol Genomics 2006, 28: 114–128
Merryweather-Clarke AT, Atzberger A, Soneji S, Gray N, Clark K, Waugh C, McGowan SJ, Taylor S, Nandi AK, Wood WG, Roberts DJ, Higgs DR, Buckle VJ, Robson KJ. Global gene expression analysis of human erythroid progenitors. Blood, 2011, 117: e96–e108
Gubin AN, Njoroge JM, Bouffard GG, Miller JL. Gene expression in proliferating human erythroid cells. Genomics 1999, 59: 168–177
Singleton BK, Burton NM, Green C, Brady RL, Anstee DJ. Mutations in EKLF/KLF1 form the molecular basis of the rare blood group in(Lu) phenotype. Blood 2008, 112: 2081–2088
Pilon AM, Arcasoy MO, Dressman HK, Vayda SE, Maksimova YD, Sangerman JI, Gallagher PG, Bodine DM. Failure of terminal erythroid differentiation in EKLF-deficient mice is associated with cell cycle perturbation and reduced expression of E2F2. Mol Cell Biol 2008, 28: 7394–7401
Kingsley PD, Greenfest-Allen E, Frame JM, Bushnell TP, Malik J, McGrath KE, Stoeckert CJ, Palis J. Ontogeny of erythroid gene expression. Blood, 2013, 121: e5–e13
Isern J, He Z, Fraser ST, Nowotschin S, Ferrer-Vaquer A, Moore R, Hadjantonakis AK, Schulz V, Tuck D, Gallagher PG, Baron MH. Single-lineage transcriptome analysis reveals key regulatory pathways in primitive erythroid progenitors in the mouse embryo. Blood 2011, 117: 4924–4934
Sripichai O, Kiefer CM, Bhanu NV, Tanno T, Noh SJ, Goh SH, Russell JE, Rognerud CL, Ou CN, Oneal PA, Meier ER, Gantt NM, Byrnes C, Lee YT, Dean A, Miller JL. Cytokine-mediated increases in fetal hemoglobin are associated with globin gene histone modification and transcription factor reprogramming. Blood 2009, 114: 2299–2306
Peller S, Tabach Y, Rotschild M, Garach-Joshua O, Cohen Y, Goldfinger N, Rotter V. Identification of gene networks associated with erythroid differentiation. Blood Cells Mol Dis 2009, 43: 74–80
An X, Schulz VP, Li J, Wu K, Liu J, Xue F, Hu J, Mohandas N, Gallagher PG. Global transcriptome analyses of human and murine terminal erythroid differentiation. Blood 2014, 123: 3466–3477
Mel HC, Prenant M, Mohandas N. Reticulocyte motility and form: studies on maturation and classification. Blood 1977, 49: 1001–1009
Come SE, Shohet SB, Robinson SH. Surface remodelling of reticulocytes produced in response to erythroid stress. Nat New Biol 1972, 236: 157–158
Chasis JA, Prenant M, Leung A, Mohandas N. Membrane assembly and remodeling during reticulocyte maturation. Blood 1989, 74: 1112–1120
Waugh RE, Mantalaris A, Bauserman RG, Hwang WC, Wu JH. Membrane instability in late-stage erythropoiesis. Blood 2001, 97: 1869–1875
Waugh RE, McKenney JB, Bauserman RG, Brooks DM, Valeri CR, Snyder LM. Surface area and volume changes during maturation of reticulocytes in the circulation of the baboon. J Lab Clin Med 1997, 129: 527–535
Liu J, Guo X, Mohandas N, Chasis JA, An X. Membrane remodeling during reticulocyte maturation. Blood 2010, 115: 2021–2027
Shi J, Kundrat L, Pishesha N, Bilate A, Theile C, Maruyama T, Dougan SK, Ploegh HL, Lodish HF. Engineered red blood cells as carriers for systemic delivery of a wide array of functional probes. Proc Natl Acad Sci USA 2014, 111: 10131–10136
Hasegawa A, Shimizu R, Mohandas N, Yamamoto M. Mature erythrocyte membrane homeostasis is compromised by loss of the GATA1-FOG1 interaction. Blood 2012, 119: 2615–2623
Moyer JD, Nowak RB, Kim NE, Larkin SK, Peters LL, Hartwig J, Kuypers FA, Fowler VM. Tropomodulin 1-null mice have a mild spherocytic elliptocytosis with appearance of tropomodulin 3 in red blood cells and disruption of the membrane skeleton. Blood 2010, 116: 2590–2599
Satchwell TJ, Bell AJ, Pellegrin S, Kupzig S, Ridgwell K, Daniels G, Anstee DJ, van den Akker E, Toye AM. Critical band 3 multiprotein complex interactions establish early during human erythropoiesis. Blood 2011, 118: 182–191
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Liu, J., Han, X. & An, X. Novel methods for studying normal and disordered erythropoiesis. Sci. China Life Sci. 58, 1270–1275 (2015). https://doi.org/10.1007/s11427-015-4971-8
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DOI: https://doi.org/10.1007/s11427-015-4971-8