Separation of Megakaryocytes From Mouse Bone Marrow by Velocity Sedimentation

Nakeff, Alexander; Maat, B.

doi:10.1182/blood.v43.4.591.591

Cited by 55 publications

(30 citation statements)

References 7 publications

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“…Platelets are formed from the cytoplasm of megakaryocytes (MKs), their precursor cells, which reside in the bone marrow (Pease, 1956). MKs are the largest (50–100 µm) and also one of the rarest cells in the bone marrow; MKs account for ∼0.01% of nucleated bone marrow cells (Nakeff and Maat, 1974). To assemble and release platelets, MKs become polyploid by endomitosis (DNA replication without cell division) and then undergo a maturation process in which the bulk of their cytoplasm is packaged into multiple long processes called proplatelets, and the nucleus is extruded.…”

Section: Introductionmentioning

confidence: 99%

The incredible journey: From megakaryocyte development to platelet formation

Machlus

Italiano

2013

Journal of Cell Biology

661

448

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Circulating blood platelets are specialized cells that prevent bleeding and minimize blood vessel injury. Large progenitor cells in the bone marrow called megakaryocytes (MKs) are the source of platelets. MKs release platelets through a series of fascinating cell biological events. During maturation, they become polyploid and accumulate massive amounts of protein and membrane. Then, in a cytoskeletal-driven process, they extend long branching processes, designated proplatelets, into sinusoidal blood vessels where they undergo fission to release platelets. Given the need for platelets in many pathological situations, understanding how this process occurs is an active area of research with important clinical applications.

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Section: Introductionmentioning

confidence: 99%

The incredible journey: From megakaryocyte development to platelet formation

Machlus

Italiano

2013

Journal of Cell Biology

661

448

View full text Add to dashboard Cite

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“…It is difficult to compare our work to others whose methods are different and whose experimental conditions and/or quantitative apprecipation of cell recovery are not very precise (Haskill et a1 1972, Nakeff & Maat 1974, Zeiller et a1 1976, Sulc et a1 1977, Wells et a1 1977a/b, Burghouts et a1 1977, 1978.…”

Section: Discussionmentioning

confidence: 95%

Separation of Human Bone Marrow Cells by Sedimentation

Guerci

Schneider

1981

Scandinavian Journal of Haematology

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Sedimentation at unit gravity of human bone marrow cells, for 15 h at 4° C on a linear density gradient of Ficoll in culture medium ranging from 1.020 to 1.065 g/ml shows that a differential migration of the bone marrow cell sub‐populations exists with precise mean densities 1.021 ±lx 10−3 g/ml for lymphocytes; 1.024 ± 2.5 times 10−3 g/ml for non‐eosinophil granulocytes; 1.025 ± 2.5 times 10−3 g/ml for metamyelocytes; 1.030 ± 3.5 times 10−3 g/ml for other myeloid cells (myeloblasts, promyelocytes, myelocytes); 1.040 ± 3 times 10−3 g/ml for eosinophil granulocytes; and 1.055 ± 10 times 10−3 g/ml for megakaryocytes. The highest percentages of S phase cell and G2 and M phase cells determined by a cytofluorograph correspond to the peaks of immature myeloid cells (myeloblasts, promyelocytes and myelocytes). This method of bone marrow cell separation may be used to study the cell cycle in pathological bone marrows (leukaemia in particular) and to determine the effects and the efficiency of some antimitotics.

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“…Several methods have been reported to be useful in the separation and enrichment of megakaryocytes and their precursors. Velocity sedimentation and centrifugal elutriation have been particularly useful for the enrichment of mature megakaryocytes because of their large size (Nakeff and Maat, 1974;Pretlow and Stinson, 1976;Worthington and Nakeff, 1981). A combination of density centrifugation in gradients of bovine serum albumin (BSA) and velocity sedimentation has made it possible to enrich for the small AchE positive cell, the earliest histochemically identifiable cell in the megakaryocytic lineage (Long et al, 1982).…”

Section: Discussionmentioning

confidence: 99%

“…AchE is an enzyme which has been shown to be relatively specific for the megakaryocytic lineage in rodents, and can be identified in small cells which are unrecognizable as megakaryocytes by standard morphologic features (Jackson, 1973;Long and Henry, 1979;Long and Williams, 1981). Techniques for the separation and partial isolation of these classes of megakaryocytes have been described (Nakeff and Maat, 1974;Leif et al, 1975; 0 1985 ALAN R. LISS, INC Levine and Fedorko, 1976;Nakeff and Floeh, 1976;Pretlow and Stinson, 1976;Nachman et al, 1977;Sitar et al, 1977;Rabellino et al, 1979;Levine, 1980;Worthington and Nakeff, 1981;Williams et al, 1981;Long et al, 1982;Sitar, 1984). Rabellino et al (1979) found that mature human megakaryocytes are found at the top of discontinuous Percoll gradients at low density (less than 1.050 g/cm3).…”

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confidence: 99%

Separation of murine megakaryocytes and their progenitors on continuous gradients of percoll

Ishibashi

Burstein

1985

Journal Cellular Physiology

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Murine bone marrow was separated on continuous Percoll density gradients to analyze the distribution of cells of the megakaryocyte lineage. Eighty-seven percent of the recovered megakaryocytes were found in fractions of density less than 1.058 g/cm3, with 63% of these cells found between 1.020 and 1.036 g/cm3. When megakaryocytes were classified according to size, 92% of the large (greater than or equal to 18 micron) acetylcholinesterase (AchE) positive cells were found in the least dense fractions (1.016-1.039 g/cm3), whereas 86% of the small (less than or equal to 10.6 micron) AchE positive cells were found in fractions of higher density (1.039-1.078 g/cm3). The distribution of enzymatic AchE activity of the separated fractions corresponded to the location of the histochemically positive cells. When ploidy measurements were made of various fractions, most of the high ploidy (32N and 64N) cells were found at low density (1.028-1.036 g/cm3), whereas no cells greater than 4N were found at density greater than 1.071 g/cm3. Thus, large AchE positive cells and the cells of highest ploidy were found at lower densities of Percoll, while small AchE positive cells and cells of low ploidy were found at higher densities. An exception to this inverse relationship was found in fractions of lowest density (less than 1.030 g/cm3) where an anomalous distribution of size and ploidy was found. The majority of megakaryocytic colony-forming cells (CFU-MK) were found at high density, as were the granulocyte-macrophage colony-forming cells (CFU-GM; approximately 1.074 g/cm3). The density distribution of the incorporation of tritiated thymidine into liquid marrow cultures was concordant with the high density distribution of colony-forming cells. The data show that megakaryocytic maturity and Percoll density varies inversely and that fractionation of marrow on continuous Percoll gradients may be a useful method for the separation and/or enrichment of megakaryocytes at different stages of differentiation.

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Separation of Megakaryocytes From Mouse Bone Marrow by Velocity Sedimentation

Cited by 55 publications

References 7 publications

The incredible journey: From megakaryocyte development to platelet formation

The incredible journey: From megakaryocyte development to platelet formation

Separation of Human Bone Marrow Cells by Sedimentation

Separation of murine megakaryocytes and their progenitors on continuous gradients of percoll

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