Secretory factors from OP9 stromal cells delay differentiation and increase the expansion potential of adult erythroid cells in vitro

Trakarnsanga, Kongtana; Wilson, Marieangela C.; Heesom, Kate J; Andrienko, Tatyana N; Srisawat, Chatchawan; Frayne, Jan

doi:10.1038/s41598-018-20491-1

Cited by 13 publications

(14 citation statements)

References 27 publications

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“…Of note, also stromal co-cultures (MS5 or OP9) do not completely result in reticulocyte maturation to erythrocytes ( Giarratana et al, 2005 ; Lu et al, 2008 ). Indeed, a recent study published a characterization of the secretory proteins from OP9, and such studies may eventually point to specific factors to will further facilitating erythropoiesis and possibly reticulocyte maturation ( Trakarnsanga et al, 2018 ). One such extrinsic signal may be transport/exchange regulation of volume control upon circulation-induced shear stress to facilitate reticulocyte maturation.…”

Section: Discussionmentioning

confidence: 99%

The Shape Shifting Story of Reticulocyte Maturation

et al. 2018

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The final steps of erythropoiesis involve unique cellular processes including enucleation and reorganization of membrane proteins and the cytoskeleton to produce biconcave erythrocytes. Surprisingly this process is still poorly understood. In vitro erythropoiesis protocols currently produce reticulocytes rather than biconcave erythrocytes. In addition, immortalized lines and iPSC-derived erythroid cell suffer from low enucleation and suboptimal final maturation potential. In light of the increasing prospect to use in vitro produced erythrocytes as (personalized) transfusion products or as therapeutic delivery agents, the mechanisms driving this last step of erythropoiesis are in dire need of resolving. Here we review the elusive last steps of reticulocyte maturation with an emphasis on protein sorting during the defining steps of reticulocyte formation during enucleation and maturation.

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Section: Discussionmentioning

confidence: 99%

The Shape Shifting Story of Reticulocyte Maturation

et al. 2018

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“…According to another report, the coculture of adult CD34 + cells with OP9 feeder cells increases the proliferation ability of erythroid cells by delaying erythroid differentiation. 35 We therefore speculated that stromal cell coculture maintains MT erythroblasts in a more immature state than the feeder-free condition, preventing their death as maturation proceeds. MT erythroblasts failed to synthesize heme and exhibited maturation defects with ring sideroblast formation in contrast to WT erythroblasts ( Figure 7 ).…”

Section: Discussionmentioning

confidence: 99%

Azacitidine is a potential therapeutic drug for pyridoxine-refractory female X-linked sideroblastic anemia

et al. 2022

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X-linked sideroblastic anemia (XLSA) is associated with mutations in the erythroid-specific δ-aminolevulinic acid synthase (ALAS2) gene. Treatment for XLSA is mainly supportive, except in pyridoxine-responsive patients. Female XLSA often represents a late onset of severe anemia, mostly due to the acquired skewing of X-chromosome inactivation. Here, we successfully generated active wild-type and mutant ALAS2 induced pluripotent stem cell (iPSC) lines from the peripheral blood cells of an affected mother and two daughters in a family with pyridoxine-resistant XLSA due to a heterozygous ALAS2 missense mutation (R227C). The erythroid differentiation potential was severely impaired in active mutant iPSC lines compared to that in active wild-type iPSC lines. Most of the active mutant iPSC-derived erythroblasts revealed an immature morphological phenotype, and some showed dysplasia and perinuclear iron deposits. Additionally, globin and HO-1 expression and heme biosynthesis in active mutant erythroblasts were severely impaired compared to that in active wild-type erythroblasts. Furthermore, genes associated with erythroblast maturation and karyopyknosis showed significantly reduced expression in active mutant erythroblasts, recapitulating the maturation defects. Notably, the erythroid differentiation ability and hemoglobin expression of active mutant iPSC-derived hematopoietic progenitor cells (HPCs) were improved by the administration of δ-aminolevulinic acid, verifying the suitability of the cells for drug testing. Administration of a DNA demethylating agent, azacitidine, reactivated the silent wild-type ALAS2 allele in active mutant HPCs and ameliorated erythroid differentiation defects, suggesting that azacitidine is a potential novel therapeutic drug for female XLSA. Our patient-specific iPSC platform provides novel biological and therapeutic insights for XLSA.

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“…Furthermore, we observed that OP9 cells impaired patient ILC2 functions, since their type-2 cytokine secretion was dramatically reduced. One can speculate that OP9-secreted factors 59 might directly affect cytokine secretion. Nevertheless, in line with previous observations, 49 we also observed that the addition of OP9 cells restrains ILC2 plasticity.…”

Section: Discussionmentioning

confidence: 99%

Healthy and Patient Type 2 Innate Lymphoid Cells are Differently Affected by in vitro Culture Conditions

Falquet¹,

Ercolano²,

Jandus³

et al. 2021

JAA

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Background Type 2 innate lymphoid cells (ILC2s) have emerged as key players in the development of type 2 driven diseases such as allergy and asthma. Due to their low number in the circulation, in vitro expansion is needed to unravel their mechanisms of action. Purpose The aim of this study is to assess the impact of different culture conditions and address whether the method of expansion may distinctly affect healthy donor or patient-derived ILC2s. Methods Here, we described the impact of six different culture conditions on the proliferation, phenotype and function of human ILC2s freshly obtained from healthy donors (healthy ILC2s) and allergic patients (patient ILC2s). Results We showed that the cytokine cocktail or the PHA induced the highest proliferation of healthy ILC2s and patient ILC2s, respectively. We observed that the stromal cells OP9, used as ILC2 feeders, did not boost their proliferation, but impaired the activation marker expression and the function of patient ILC2s. Furthermore, we demonstrated that the culture conditions differently impacted the activation state of c-Kit high and c-Kit low ILC2s, in both healthy donors and allergic patients. Last, we also observed that ILC2s expanded only with IL-2 and IL-7 were the most prone to secrete IL-5 and IL-13 upon IL-33 stimulation. In contrast, in patients, the addition of OP9 cells during the expansion restrained their type 2 cytokine secretory functions. Conclusion This report highlights that culture conditions distinctly impacted on the healthy or patient ILC2 behavior, with important consequences for their study in disease settings.

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Secretory factors from OP9 stromal cells delay differentiation and increase the expansion potential of adult erythroid cells in vitro

Cited by 13 publications

References 27 publications

The Shape Shifting Story of Reticulocyte Maturation

The Shape Shifting Story of Reticulocyte Maturation

Azacitidine is a potential therapeutic drug for pyridoxine-refractory female X-linked sideroblastic anemia

Healthy and Patient Type 2 Innate Lymphoid Cells are Differently Affected by in vitro Culture Conditions

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