Acute megakaryoblastic leukemia is a rare and rapidly fatal disease characterized by proliferation of megakaryocyte series and atypical megakaryocytes in the bone marrow. Acute megakaryoblastic leukemia is suspicious when 1) megakaryocyte in peripheral blood, mixture of large and small mononuclear megakaryoblast in the bone marrow 2) cytoplasmic budding in blast 3) myelofibrosis (dense medullary overgrowth of reticulin fibers) 4) PAS (+), ANAE (+), SBB (−), peroxidase (−) and which is confirmed by platelet peroxidase oxidation on electron microscope or monoclonal antibody. A case of acute megakaryoblastic leukemia was studied morphologically and monoclonal antibody.
Blood Platelets ; Bone Marrow ; Cytoplasm ; Leukemia, Megakaryoblastic, Acute* ; Megakaryocyte Progenitor Cells ; Megakaryocytes ; Naphthol AS D Esterase ; Peroxidase ; Primary Myelofibrosis ; Reticulin

OBJECTIVE: The purposes of this study were to isolate and characterize mesenchymal stem cells (MSCs) with osteogenic and adipogenic potential from umbilical cord blood (UCB). METHODS: MSCs were isolated using a density gradient centrifugation and extensive subcultivation from UCB. The proliferation capability, cell cycle, cytochemical markers, and immunophenotype of these MSCs were measured. The transcript of osteoblast-specific markers, alkaline phosphatase (ALP), and calcium deposit were analyzed from MSCs cultured in osteogenic media for 1-4 weeks, and MSCs exposed in adipogenic media were stained by Oil red after 4 weeks of culture. RESULTS: More than 84% of MSCs were in the G0/G1 phase of cell cycle. MSCs were positive for periodic acid-Schiff, and alpha-naphthyl acetate esterase activity, but negative for sudan black-B, and ALP activities. MSCs expressed CD13, CD29, CD 44, CD49e, CD51, CD54, CD 90, SH2, and SH3, but did not express CD11b, CD14, CD31, CD34, CD45, CD49d, CD106, CD133, CD144, CD146, CD163, CD166, and Stro-1. When MSCs were cultured in osteogenic media for 1-4 weeks, they expressed transcripts of osteoblastic specific markers: Runx-2, ALP, and procollagen type I. During culture of 2-4 weeks, ALP activity was detected and quantification increased significantly, and the deposition of a calcified matrix became evident. Exposure of MSCs to adipogenic media resulted in morphological change stained by Oil Red O. CONCLUSION: These data indicate that UCB contains MSCs with osteogenic and adipogenic differentiation potential, and may serve as an potential source of MSCs to be utilized in cell therapy for various diseases.
Alkaline Phosphatase ; Calcium ; Cell Cycle ; Cell- and Tissue-Based Therapy ; Centrifugation, Density Gradient ; Collagen Type I ; Fetal Blood* ; Mesenchymal Stromal Cells* ; Naphthol AS D Esterase ; Osteoblasts ; Sudan ; Umbilical Cord*

OBJECTIVE: The purposes of this study were to isolate and characterize mesenchymal stem cells (MSCs) with osteogenic and adipogenic potential from umbilical cord blood (UCB). METHODS: MSCs were isolated using a density gradient centrifugation and extensive subcultivation from UCB. The proliferation capability, cell cycle, cytochemical markers, and immunophenotype of these MSCs were measured. The transcript of osteoblast-specific markers, alkaline phosphatase (ALP), and calcium deposit were analyzed from MSCs cultured in osteogenic media for 1-4 weeks, and MSCs exposed in adipogenic media were stained by Oil red after 4 weeks of culture. RESULTS: More than 84% of MSCs were in the G0/G1 phase of cell cycle. MSCs were positive for periodic acid-Schiff, and alpha-naphthyl acetate esterase activity, but negative for sudan black-B, and ALP activities. MSCs expressed CD13, CD29, CD 44, CD49e, CD51, CD54, CD 90, SH2, and SH3, but did not express CD11b, CD14, CD31, CD34, CD45, CD49d, CD106, CD133, CD144, CD146, CD163, CD166, and Stro-1. When MSCs were cultured in osteogenic media for 1-4 weeks, they expressed transcripts of osteoblastic specific markers: Runx-2, ALP, and procollagen type I. During culture of 2-4 weeks, ALP activity was detected and quantification increased significantly, and the deposition of a calcified matrix became evident. Exposure of MSCs to adipogenic media resulted in morphological change stained by Oil Red O. CONCLUSION: These data indicate that UCB contains MSCs with osteogenic and adipogenic differentiation potential, and may serve as an potential source of MSCs to be utilized in cell therapy for various diseases.
Alkaline Phosphatase ; Calcium ; Cell Cycle ; Cell- and Tissue-Based Therapy ; Centrifugation, Density Gradient ; Collagen Type I ; Fetal Blood* ; Mesenchymal Stromal Cells* ; Naphthol AS D Esterase ; Osteoblasts ; Sudan ; Umbilical Cord*

BACKGROUND: In general, internal/external quality control of special stains for diagnosis of hematological diseases may be unavailable in a clinical laboratory owing to the lack of an appropriate positive/negative control material. METHODS: We developed a protocol on positive/negative control materials for five special stains (iron, myeloperoxidase [MPO], periodic acid-Schiff [PAS], Sudan black B [SBB], and alpha-naphthyl acetate esterase [ANAE]) using a hematological malignant cell line. First, we compared stainability of seven cell lines (HL-60, THP-1, K562, Kasumi-1, KG-1, KO52, and NKM-1), then confirmed duration of stable stainability. A proficiency test using external quality control materials was conducted at eleven institutions, which participated voluntarily. RESULTS: HL-60 and THP-1 cell lines, which showed good stainability among the seven cancer cell lines, were selected as external quality control materials. The stainability of a prepared cell line fixed on control slides was stable for 3–4 weeks (MPO, SBB, and PAS) or 9–10 weeks (ANAE). The stainability of paraffin-embedded control material for iron stain was stable for 3 months. The results from 11 institutions were the same on iron, MPO, SBB, and ANAE. Nevertheless, two of 10 institutes showed discrepant results on PAS. CONCLUSIONS: In this study, we demonstrated that cell lines could serve as a standard quality control material for special stains. Most institutions showed representative results on special stains except for PAS. This protocol for special stain may be useful as an external or internal quality control in a haematology laboratory.
Academies and Institutes ; Cell Line ; Coloring Agents* ; Diagnosis ; Hematologic Diseases ; Hematology* ; Iron ; Laboratory Proficiency Testing ; Naphthol AS D Esterase ; Peroxidase ; Quality Control* ; Sudan