1994
DOI: 10.1007/bf00157966
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Immunocytochemical identification of murine and human megakaryocyte colonies in soft-agar cultures

Abstract: Murine megakaryocyte (MK) colonies in soft-agar cultures were immunocytochemically stained with platelet antiserum and an immuno-alkaline phosphatase procedure. Subsequently, cytochemical staining for acetylcholinesterase was used to confirm the specificity of the immunolabelling technique. The correlation of numbers of megakaryocyte colonies enumerated by independent observers was excellent. A comparable platelet antiserum directed against human platelet epitopes was utilized to identify human MK colonies in … Show more

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Cited by 5 publications
(4 citation statements)
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“…We therefore established and validated an immunoenzyme-based method of staining monoclonal antibody-labeled human megakaryocyte colonies in situ. This method is relatively permanent and less time consuming than fluorescent staining approaches, while offering excellent maintenance of colony morphology and, in our hands, is substantially more robust than identification in plasma clot cultures [11,[33][34][35]. 38 Mpl Ligand Prolongs Progenitor Cell Survival Before embarking upon studies of megakaryocyte survival, it was important to establish the linearity of response of megakaryocyte precursor cells in our assay.…”
Section: Discussionmentioning
confidence: 99%
See 1 more Smart Citation
“…We therefore established and validated an immunoenzyme-based method of staining monoclonal antibody-labeled human megakaryocyte colonies in situ. This method is relatively permanent and less time consuming than fluorescent staining approaches, while offering excellent maintenance of colony morphology and, in our hands, is substantially more robust than identification in plasma clot cultures [11,[33][34][35]. 38 Mpl Ligand Prolongs Progenitor Cell Survival Before embarking upon studies of megakaryocyte survival, it was important to establish the linearity of response of megakaryocyte precursor cells in our assay.…”
Section: Discussionmentioning
confidence: 99%
“…We therefore established and validated an immunoenzyme‐based method of staining monoclonal antibody‐labeled human megakaryocyte colonies in situ. This method is relatively permanent and less time consuming than fluorescent staining approaches, while offering excellent maintenance of colony morphology and, in our hands, is substantially more robust than identification in plasma clot cultures [11, 33‐35].…”
Section: Discussionmentioning
confidence: 99%
“…CFU-M and CFU-G were scored by morphology, whereas CFU-Meg were identified by histochemical staining for acetylcholinesterase. Colonies containing at least three acetylcholinesterase-positive cells were scored as CFU-Meg (30).…”
Section: Methodsmentioning
confidence: 99%
“…31 Colonies containing at least 3 acetylcholinesterase-positive cells were scored as colony-forming unit-megakaryocyte (CFU-MK). 32 For megakaryocyte cultures, femurs and tibias were flushed with CATCH medium, and cells were subjected to lineage depletion (Mouse Hematopoietic Cell Lineage Depletion Kit; R&D Systems), leading to a relative enrichment of megakaryocytes. RNA was prepared from cells immediately or after 2 or 4 days of culture in RPMI with 10% fetal calf serum (FCS), Tpo-conditioned media, 6 and 10 g/mL recombinant murine stem cell factor (BioSource, Camarillo, CA).…”
Section: Hematopoietic Progenitors Megakaryocyte Culture and Ploidymentioning
confidence: 99%