In Vitro Mass Production of Human Erythroid Cells from the Blood of Normal Donors and of Thalassemic Patients

Migliaccio, Giovanni; Pietro, Roberta Di; Giacomo, Viviana di; Baldassarre, Angela Di; Migliaccio, Anna Rita; Maccioni, Liliana; Galanello, Renzo; Papayannopoulou, Thalia

doi:10.1006/bcmd.2002.0502

Cited by 130 publications

(175 citation statements)

References 16 publications

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“…CD36 is typically found on erythroid and megakaryocytic progenitor cells, but it appears earlier on the cells of erythroid lineage and has been defined as a marker for EPCs (6,20). During maturation to erythroblasts, cells also express CD71 (12) and GPA (12). An analysis of the cell surface antigens of the CD34 ϩ cells selected from PBSCs indicated an absence of CD36 and GPA.…”

Section: Discussionmentioning

confidence: 99%

Ex Vivo-Generated CD36 ⁺ Erythroid Progenitors Are Highly Permissive to Human Parvovirus B19 Replication

Wong

Zhi

Filippone

et al. 2008

J Virol

119

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The pathogenic parvovirus B19 (B19V) has an extreme tropism for human erythroid progenitor cells. In vitro, only a few erythroid leukemic cell lines (JK-1 and KU812Ep6) or megakaryoblastoid cell lines (UT7/Epo and UT7/Epo-S1) with erythroid characteristics support B19V replication, but these cells are only semipermissive. By using recent advances in generating large numbers of human erythroid progenitor cells (EPCs) ex vivo from hematopoietic stem cells (HSCs), we produced a pure population of CD36 ؉ EPCs expanded and differentiated from CD34 ؉ HSCs and assessed the CD36 ؉ EPCs for their permissiveness to B19V infection. Over more than 3 weeks, cells grown in serum-free medium expanded more than 800,000-fold, and 87 to 96% of the CD36 ؉ EPCs were positive for globoside, the cellular receptor for B19V. Immunofluorescence (IF) staining showed that about 77% of the CD36 ؉ EPCs were positive for B19V infection, while about 9% of UT7/Epo-S1 cells were B19V positive. Viral DNA detected by real-time PCR increased by more than 3 logs in CD36 ؉ EPCs; the increase was 1 log in UT7/Epo-S1 cells. Due to the extensive permissivity of CD36 ؉ EPCs, we significantly improved the sensitivity of detection of infectious B19V by real-time reverse transcription-PCR and IF staining 100-and 1,000-fold, respectively, which is greater than the sensitivity of UT7/Epo-S1 cell-based methods. This is the first description of an ex vivo method to produce large numbers of EPCs that are highly permissive to B19V infection and replication, offering a cellular system that mimics in vivo infection with this pathogenic human virus.

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Section: Discussionmentioning

confidence: 99%

Ex Vivo-Generated CD36 ⁺ Erythroid Progenitors Are Highly Permissive to Human Parvovirus B19 Replication

Wong

Zhi

Filippone

et al. 2008

J Virol

119

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“…The finding that p57 KIP2 is an essential mediator of Dex-dependent CFUe expansion cultures provides a novel insight into the mechanism by which glucocorticoids sustain these cultures, and is of potential translational relevance in the development of mass production of erythroid progenitors in vitro that rely on this culture system (52,80).…”

Section: Cip1mentioning

confidence: 99%

Global increase in replication fork speed during a p57KIP2-regulated erythroid cell fate switch

Socolovsky

Futran

Hidalgo

2017

Experimental Hematology

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Cell cycle regulators are increasingly implicated in cell fate decisions, such as the acquisition or loss of pluripotency and self-renewal potential. The cell cycle mechanisms that regulate these cell fate decisions are largely unknown. We studied an S phase-dependent cell fate switch, in which murine early erythroid progenitors transition in vivo from a self-renewal state into a phase of active erythroid gene transcription and concurrent maturational cell divisions. We found that progenitors are dependent on p57 KIP2 -mediated slowing of replication forks for self-renewal, a novel function for cyclin-dependent kinase inhibitors. The switch to differentiation entails rapid down-regulation of p57 KIP2 with a consequent global increase in replication fork speed and an abruptly shorter S phase. Our work suggests that cell cycles with specialized global DNA replication dynamics are integral to the maintenance of specific cell states and to cell fate decisions.

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“…They have been shown to induce apoptosis in lymphoid cells [16], prevent apoptosis in granulocytes [17], and drive the proliferation of erythroid progenitors [18,19].…”

Section: Molecular Mechanism Of Glucocorticoidsmentioning

confidence: 99%

Functional Links Between Glucocorticoids and Cytokines In DBA

2015

J Paediatr Child Care

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Diamond Blackfan anaemia (DBA) is a red blood cell aplasia characterized by erythropoietic defects as well as congenital anomalies. Forty percent of patients with DBA are treated with glucocorticoid steroids, which remain the mainstay of treatment in DBA. Many advances in the understanding of the physiological role of the glucocorticoid receptor have been made since the first introduction of glucocorticoids to the clinic, but their mechanism of action in the treatment of DBA is still under investigation. This review is intended to summarize the mechanism of glucocorticoid action specifically as related to erythropoiesis, focusing on the functional links between glucocorticoids and cytokines. IntroductionDiamond Blackfan anemia (DBA) was first described as a disorder of impaired red blood cell production in children [1,2]. While most of the DBA cases are diagnosed in early infancy, a recent case report reveals that DBA can occur during fetal development. This suggests that severely affected DBA fetus likely die to hydrops fetalis and result in undiagnosed miscarriages [3]. This disorder results from a cellular defect in which erythroid progenitors are highly sensitive to death by apoptosis, leading to erythropoietic failure [4]. The etiology of DBA has been the subject of continuous discussions, and while the early success of treatment with glucocorticoids (GCs) in 1951 [5] led to the idea that the pathogenesis of DBA could have an immunologic basis [6], it is now accepted that DBA is in fact a member of a rare group of genetic disorders.Approximately 50% of DBA patients have a single mutation in a gene encoding a ribosomal protein, which indicates that DBA is associated with a disorder of ribosome biogenesis and/or function [4]. The pronounced erythroid defect suggests that erythroid progenitors may express specific mRNAs that are hypersensitive to the decreased translation capacity [7]. Twenty-five percent of patients have a mutation in the ribosomal protein S19 (RPS19) gene [8], and two independent studies have demonstrated that over expression of the RPS19 transgene increases the number of erythroid colonies in RPS19 deficient hematopoietic progenitor cells in vitro [9,10]. In a mouse model of DBA, a high expression of RPS19 can rescue the erythroid development, and the corrected DBA cells show a survival advantage in vivo [11]. As zebrafish hematopoietic regulation is conserved with mammals, zebrafish models have also been reported to be useful in studying DBA [12][13][14], recapitulating many aspects of the DBA phenotype, including hematopoietic specific defects and p53 activation [14]. The list of genes that are mutated in DBA has been updated in 2013, to include ten ribosomal genes and one transcriptional regulator: RPS19, RPS24, RPS17, RPL35A, RPL5, RPL11, RPS7, RPS10, RPS26, RPL26, and GATA1 [15].More than 50 years after their introduction to the clinic, GCs are still the most effective drugs used in DBA. Nevertheless, the reported side effects of GCs include decreased growth velocity in infants...

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In Vitro Mass Production of Human Erythroid Cells from the Blood of Normal Donors and of Thalassemic Patients

Cited by 130 publications

References 16 publications

Ex Vivo-Generated CD36 ⁺ Erythroid Progenitors Are Highly Permissive to Human Parvovirus B19 Replication

Ex Vivo-Generated CD36 ⁺ Erythroid Progenitors Are Highly Permissive to Human Parvovirus B19 Replication

Global increase in replication fork speed during a p57KIP2-regulated erythroid cell fate switch

Functional Links Between Glucocorticoids and Cytokines In DBA

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In Vitro Mass Production of Human Erythroid Cells from the Blood of Normal Donors and of Thalassemic Patients

Cited by 130 publications

References 16 publications

Ex Vivo-Generated CD36 + Erythroid Progenitors Are Highly Permissive to Human Parvovirus B19 Replication

Ex Vivo-Generated CD36 + Erythroid Progenitors Are Highly Permissive to Human Parvovirus B19 Replication

Global increase in replication fork speed during a p57KIP2-regulated erythroid cell fate switch

Functional Links Between Glucocorticoids and Cytokines In DBA

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Ex Vivo-Generated CD36 ⁺ Erythroid Progenitors Are Highly Permissive to Human Parvovirus B19 Replication

Ex Vivo-Generated CD36 ⁺ Erythroid Progenitors Are Highly Permissive to Human Parvovirus B19 Replication