Improved Plasma Culture System for Production of Erythrocytic Colonies In Vitro: Quantitative Assay Method for CFU-E

McLeod, Donald W.; Shreeve, Mona M.; Axelrad, Arthur A.

doi:10.1182/blood.v44.4.517.517

Cited by 465 publications

(70 citation statements)

References 5 publications

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“…The EEC was assessed in vitro, in the absence of exogenous erythropoietin, in a single laboratory. The EEC was assessed in 118 patients, using collected BM mononuclear cells, as previously described (McLeod et al, 1974). We analysed the results for 104 patients whose control cultures displayed normal colony growth in the presence of exogenous erythropoietin.…”

Section: Methodsmentioning

confidence: 99%

Relevance of the criteria commonly used to diagnose myeloproliferative disorder in patients with splanchnic vein thrombosis

Condat²,

et al. 2005

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Summary Myeloproliferative disorders (MPD) are reported in 25–65% of patients with splanchnic vein thrombosis (SVT). Diagnostic criteria for MPD have not been fully established in this context. Using clusters of abnormal megakaryocytes in bone marrow (BM) biopsy as a reference standard for Philadelphia negative MPD, we assessed the relevance of other criteria currently recommended for the diagnosis of MPD in SVT (128 consecutive SVT patients). First, usual criteria were compared with BM results: endogenous erythroid colony formation (EEC) was strongly correlated with BM results; splenomegaly, blood cell count, total red cell volume, erythropoietin level and cytogenetic were much less accurate. Then, patients were assigned to three groups according to the combination of BM and EEC findings (group I: both present; group II: both absent; group III: other patients); clinical presentation and outcome were compared in each group. At a mean follow‐up of 6·09 ± 6·6 years, progression to a severe form of MPD occurred in 7 of 31 group I patients (23%), in 1 of 34 group III patients (3%) and 0 of 63 group II patients. The combination of marked splenomegaly and platelet count >200 × 109/l was restricted to groups I and III. In conclusion, in patients with SVT, BM findings and EEC allowed the diagnosis of MPD at risk of aggravation. Marked splenomegaly in association with platelet counts >200 × 109/l constitute a simple index with high specificity but low sensitivity.

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Section: Methodsmentioning

confidence: 99%

Relevance of the criteria commonly used to diagnose myeloproliferative disorder in patients with splanchnic vein thrombosis

Condat²,

et al. 2005

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“…A number ofstudies have demonstrated that Ep is essential for EC formation in bone marrow cultures (Iscove ct a/, 1974;McLeod et a / , 1974;Stephenson et al, 1971). However, the interaction of Ep ivith CFU-E has not been clearly defined since Ep is usually present throughout the culture period.…”

Section: Discussionmentioning

confidence: 99%

Effects of Shortened Erythropoietin Exposure on Sheep Marrow Cultures

Roodman

Kaplan

et al. 1981

Br J Haematol

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To determine whether erythropoietin (Ep) is required only to trigger, or also to sustain the growth of erythroid progenitor (CFU-E) in vitro, the effect of limiting the time of exposure of sheep bone marrow CFU-E to Ep on erythroid colony (EC) formation was examined. Restricting the in vitro exposure of CFU-E to 2-24 h, either by brief preincubation with Ep or by neutralization of Ep present in culture with anti-Ep, resulted in a 70-90% reduction of EC formation, and the colonies that were produced were of a small size. In contrast, no inhibition of EC size or number occurred when CFU-E were exposed to EP for more than 24 h. These results suggest that Ep also modulates the proliferative events subsequent to the initial triggering of CFU-E which lead to the formation of erythrocytes.

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“…The mononuclear cells were isolated by Ficoll-metrizoate gradient density centrifugation. The collected cells were washed three times in a a medium and were subsequently plated a t a concentration of 10" in 3 5 m m petri dishes (Falcon) in a I ml volume by the plasma clot technique (McLeod et al, 1974;Clarke & Housman, 1977). This technique has been slightly modified: (i) a medium was used instead of N C T C 109 and M.E.M.…”

Section: Materials a N D Methodsmentioning

confidence: 99%

Morphological Abnormalities in Cultured Erythroid Colonies (BFU‐E) from the Blood of Two Patients with HEMPAS

1979

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The results of cytological and ultrastructural analysis of erythroid burst colonies derived from the peripheral blood of two patients with HEMPAS have been compared to those obtained in normal controls. Using the plasma clot technique, in studies on 10 subjects we confirmed that most of the colonies consisted or erythroblasts with a synchronous and normal maturation involving a wave of nuclear extrusion at day 13. In contrast, the majority of well-haemoglobinized colonies from HEMPAS consisted of numerous bi- or multinucleated erythroblasts displaying the supplementary double membrane beneath their plasma membrane. This excessive membrane may be present as a continuous or fragmented structure in different erythroblasts from the same colony. These findings suggest that the progeny derived from one BFU-E may vary considerably in their morphological defects. Furthermore, one third of the packed colonies appeared to be formed by non-haemoglobinized cells which were clearly identified by electron microscopy as very early erythroblasts. These cells were unable to mature and subsequently lysed. Thus dyserythropoiesis occurred in culture both at early and late stages of maturation. These studies clearly demonstrate that HEMPAS is a disorder resulting from defective erythroid committed cells.

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Improved Plasma Culture System for Production of Erythrocytic Colonies In Vitro: Quantitative Assay Method for CFU-E

Cited by 465 publications

References 5 publications

Relevance of the criteria commonly used to diagnose myeloproliferative disorder in patients with splanchnic vein thrombosis

Relevance of the criteria commonly used to diagnose myeloproliferative disorder in patients with splanchnic vein thrombosis

Effects of Shortened Erythropoietin Exposure on Sheep Marrow Cultures

Morphological Abnormalities in Cultured Erythroid Colonies (BFU‐E) from the Blood of Two Patients with HEMPAS

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