Establishment of an erythroid cell line from primary CD36+ erythroid progenitor cells

Wong, Susan; Keyvanfar, Keyvan; Wan, Zhihong; Young, Neal S.; Zhi, Ning

doi:10.1016/j.exphem.2010.07.012

Cited by 17 publications

(16 citation statements)

References 54 publications

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“…However, deficiency of CD36 has no effect on development of erythroblasts (Toba et al., 2001). CD36, in our present study, is only an effective co-marker for tracing the developing erythroid lineage cells, a method that has been widely used (Wong et al., 2010, Okumura et al., 1992). …”

Section: Discussionmentioning

confidence: 86%

Early Development of Definitive Erythroblasts from Human Pluripotent Stem Cells Defined by Expression of Glycophorin A/CD235a, CD34, and CD36

Mao

Huang

et al. 2016

Stem Cell Reports

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SummaryThe development of human erythroid cells has been mostly examined in models of adult hematopoiesis, while their early derivation during embryonic and fetal stages is largely unknown. We observed the development and maturation of erythroblasts derived from human pluripotent stem cells (hPSCs) by an efficient co-culture system. These hPSC-derived early erythroblasts initially showed definitive characteristics with a glycophorin A+ (GPA+) CD34lowCD36− phenotype and were distinct from adult CD34+ cell-derived ones. After losing CD34 expression, early GPA+CD36− erythroblasts matured into GPA+CD36low/+ stage as the latter expressed higher levels of β-globin along with a gradual loss of mesodermal and endothelial properties, and terminally suppressed CD36. We establish a unique in vitro model to trace the early development of hPSC-derived erythroblasts by serial expression of CD34, GPA, and CD36. Our findings may provide insight into the understanding of human early erythropoiesis and, ultimately, therapeutic potential.

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Section: Discussionmentioning

confidence: 86%

Early Development of Definitive Erythroblasts from Human Pluripotent Stem Cells Defined by Expression of Glycophorin A/CD235a, CD34, and CD36

Mao

Huang

et al. 2016

Stem Cell Reports

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“…To our knowledge, this is the first report describing the establishment of immortalized human erythroid progenitor cell lines able to produce enucleated RBCs ex vivo. Although erythroid progenitor cell lines have previously been established using HPV16-E6/E7, these cell lines did not produce enucleated RBCs [17]. One possible explanation for this failure is the continuous expression of HPV16-E6/E7 in these cells.…”

Section: Discussionmentioning

confidence: 95%

“…Enforced expression of human papilloma virus (HPV) 16‐E6/E7 protein in CD36‐positive erythroid cells has recently been used to generate immortalized cell lines [7]. We therefore decided to use this approach with human umbilical cord blood cells and iPS cells as the source cell materials.…”

Section: Can Immortalized Human Erythroid Progenitor Cell Lines Be Prmentioning

confidence: 99%

“…Immortalized erythroid progenitor cell lines can be established by transformation of erythroid progenitor cells with the HPV16-derived proteins HPV16-E6/E7 [17]. These cell lines do not produce mature erythrocytes, such as enucleated RBCs, possibly because continuous expression of HPV16-E6/E7 inhibits terminal differentiation of the cells.…”

Section: Establishment Of Immortalized Erythroid Progenitor Cell Linementioning

confidence: 99%

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Establishment of immortalized human erythroid progenitor cell lines able to produce enucleated red blood cells

Nakamura

2012

ISBT Science Series

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Transfusion of red blood cells (RBCs) is a standard and indispensable therapy in current clinical practice. In vitro production of RBCs offers a potential means to overcome a shortage of transfusable RBCs in some clinical situations and also to provide a source of cells free from possible infection or contamination by microorganisms. Thus, in vitro production of RBCs may become a standard procedure in the future. We previously reported the successful establishment of immortalized mouse erythroid progenitor cell lines that were able to produce mature RBCs very efficiently. Here, we have developed a reliable protocol for establishing immortalized human erythroid progenitor cell lines that are able to produce enucleated RBCs. These immortalized cell lines produce functional hemoglobin and express erythroid-specific markers, and these markers are upregulated following induction of differentiation in vitro. Most importantly, these immortalized cell lines all produce enucleated RBCs after induction of differentiation in vitro, although the efficiency of producing enucleated RBCs remains to be improved further. To the best of our knowledge, this is the first demonstration of the feasibility of using immortalized human erythroid progenitor cell lines as an ex vivo source for production of enucleated RBCs.

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“…15 Most of these genetically immortalized erythroid cell lines are of leukemic cell origin or transformed by in vitro genetic manipulation, and thus have defects on terminal differentiation and maturation, rendering them unsuitable for clinical application. [16][17][18] Recently, mouse erythroblast lines have been established from in vitro differentiated ESCs or early mouse embryos that have normal in vitro or in vivo terminal maturation capabilities. [19][20][21] These new findings suggest that embryonic stage erythroblasts process much higher proliferative or self-renewal capabilities than postnatal counterparts.…”

Section: Introductionmentioning

confidence: 99%

Extensive Ex Vivo Expansion of Functional Human Erythroid Precursors Established From Umbilical Cord Blood Cells by Defined Factors

et al. 2014

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There is a constant shortage of red blood cells (RBCs) from sufficiently matched donors for patients who need chronic transfusion. Ex vivo expansion and maturation of human erythroid precursors (erythroblasts) from the patients or optimally matched donors could represent a potential solution. Proliferating erythroblasts can be expanded from umbilical cord blood mononuclear cells (CB MNCs) ex vivo for 10(6)-10(7)-fold (in ~50 days) before proliferation arrest and reaching sufficient number for broad application. Here, we report that ectopic expression of three genetic factors (Sox2, c-Myc, and an shRNA against TP53 gene) associated with iPSC derivation enables CB-derived erythroblasts to undergo extended expansion (~10(68)-fold in ~12 months) in a serum-free culture condition without change of cell identity or function. These expanding erythroblasts maintain immature erythroblast phenotypes and morphology, a normal diploid karyotype and dependence on a specific combination of growth factors for proliferation throughout expansion period. When being switched to a terminal differentiation condition, these immortalized erythroblasts gradually exit cell cycle, decrease cell size, accumulate hemoglobin, condense nuclei and eventually give rise to enucleated hemoglobin-containing erythrocytes that can bind and release oxygen. Our result may ultimately lead to an alternative approach to generate unlimited numbers of RBCs for personalized transfusion medicine.

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Establishment of an erythroid cell line from primary CD36+ erythroid progenitor cells

Abstract: The continuous CD36E cell line is an erythroid progenitor cell line possessing the ability to produce hemoglobin. The CD36E cell line would be an excellent tool for applied research involving erythroid lineage cells and comparative studies with primary CD36(+) EPCs.

Cited by 17 publications

References 54 publications

Early Development of Definitive Erythroblasts from Human Pluripotent Stem Cells Defined by Expression of Glycophorin A/CD235a, CD34, and CD36

Early Development of Definitive Erythroblasts from Human Pluripotent Stem Cells Defined by Expression of Glycophorin A/CD235a, CD34, and CD36

Establishment of immortalized human erythroid progenitor cell lines able to produce enucleated red blood cells

Extensive Ex Vivo Expansion of Functional Human Erythroid Precursors Established From Umbilical Cord Blood Cells by Defined Factors

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