Cold synchronization for the study of peripheral blood and bone marrow chromosomes in leukemias and other hematologic disease states

Boucher, B. J.; Norman, C. S.

doi:10.1007/bf00278974

Cited by 23 publications

(2 citation statements)

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“…Cytogenetic Analysis: Karyotypes were obtained from bone marrow aspirates, peripheral blood, and established cell cultures using a variety of synchronization and banding techniques (14)(15)(16). Most cells showing aneuploidy, polyploidy, or structural anomalies were karyotyped to assess the extent and nature of the anomalies.…”

Section: Cytocentrifuge Preparations: Fresh Blood Mononuclear Cells Ob-mentioning

confidence: 99%

Novel peripheral blood-derived human cell lines with properties of megakaryocytes.

Morgan¹,

Brodsky²

1985

The Journal of Cell Biology

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For 18 mo, we derived 18 cell lines from 11 donors with various clinical profiles ranging from normal to leukemic. Suspension cultures were initiated with 1 x 106 mononuclear blood cells/ml of nutrient medium containing 10% human serum and 10% lectin-stimulated human lymphocyte conditioned medium. The cultures were monitored weekly by morphological analyses of Wright-Giemsa-stained cell preparations. All successful cultures showed a significant decline in viability during the first 3-4 wk with rare "lymphoid" cells observed in mitosis. Within the next 2 wk, the proliferating cells gave rise to a rapidly expanding population of mononuclear cells. As the cultures expanded, cell morphology became heterogeneous with respect to cell size and nuclear ploidy, with an accumulation of giant multinuclear cells that were suggestive of megakaryocytes. Even though the cells did not have the classical morphology of mature platelet-forming megakaryocytes, 90% of the cells within a cell line were positive by direct or indirect immunofluorescence for the platelet membrane glycoproteins lib and Ilia; for surface markers HLA-Dr and B2-microglobulin; for intracellular plateletderived growth factor and platelet factor IV; and for membrane affinity or binding with serum platelet-derived growth factor and platelet factor IV. These results suggest that a blood precursor cell, most likely a primitive megakaryoblast, was isolated from the peripheral blood and was provided with an optimal culture environment for sustained growth. These cells did not mature to a more differentiated stage, perhaps owing to regulatory factor deficiencies in this in vitro system. The remarkable frequency of obtaining cell lines with megakaryocyte properties from normal peripheral blood and the capacity of some normal donors to repeatedly yield these cell lines make this cell culture system indeed unique by being selective for putative megakaryocyte precursors.

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Section: Cytocentrifuge Preparations: Fresh Blood Mononuclear Cells Ob-mentioning

confidence: 99%

Novel peripheral blood-derived human cell lines with properties of megakaryocytes.

Morgan¹,

Brodsky²

1985

The Journal of Cell Biology

View full text Add to dashboard Cite

show abstract

“…As respostas celulares ao frio, além de alterações na progressão do ciclo, abrangem desnaturação e desarranjo de proteínas, inibição de transcrição e tradução, alterações nos elementos do citoesqueleto e mudanças na permeabilidade da membrana plasmática (SONNA et al, 2002). Boucher & Norman (1980) Alguns autores propõem o uso de bloqueio duplo para obtenção de células em diversas fases do ciclo (UZBEKOV et al, 1998). No entanto, os métodos devem ser padronizados para cada tipo celular, considerando a variabilidade das respostas de cada linhagem ao tratamento utilizado e a duração de cada fase do ciclo.…”

Section: Discussionunclassified