scholarly journals The proportion of mitoses in different cell lineages changes during short-term culture of normal human bone marrow

Blood ◽  
1986 ◽  
Vol 67 (5) ◽  
pp. 1240-1243 ◽  
Author(s):  
M Keinanen ◽  
S Knuutila ◽  
CD Bloomfield ◽  
E Elonen ◽  
A de la Chapelle

Abstract To determine the hematopoietic cell lineage of mitotic cells in human bone marrow on direct examination and after 24-hour culture, marrow mitoses from four healthy individuals were studied, using a new technique that allows analysis of karyotypes in cells whose cell membrane and cytoplasm have been preserved. Mitoses were identified as being of erythroid lineage by immunofluorescent staining for surface glycophorin A and as being of granulocytic lineage by cytoplasmic staining for Sudan black B. On direct marrow examination without prior culture, the great majority of mitoses (74% to 90%) were of erythroid lineage; only a few (0% to 10%) were granulocytic. After 24-hour culture, the percentage of erythroid mitoses (15% to 40%) decreased, while the percentage of granulocytic mitoses (58% to 87%) increased strikingly. These data indicate that mitotic cells of different hematopoietic cell lineages predominate in marrow at different culture times and offer a plausible explanation for the high frequency of normal karyotypes in acute myeloid leukemia after direct marrow cytogenetic evaluation.

Blood ◽  
1986 ◽  
Vol 67 (5) ◽  
pp. 1240-1243
Author(s):  
M Keinanen ◽  
S Knuutila ◽  
CD Bloomfield ◽  
E Elonen ◽  
A de la Chapelle

To determine the hematopoietic cell lineage of mitotic cells in human bone marrow on direct examination and after 24-hour culture, marrow mitoses from four healthy individuals were studied, using a new technique that allows analysis of karyotypes in cells whose cell membrane and cytoplasm have been preserved. Mitoses were identified as being of erythroid lineage by immunofluorescent staining for surface glycophorin A and as being of granulocytic lineage by cytoplasmic staining for Sudan black B. On direct marrow examination without prior culture, the great majority of mitoses (74% to 90%) were of erythroid lineage; only a few (0% to 10%) were granulocytic. After 24-hour culture, the percentage of erythroid mitoses (15% to 40%) decreased, while the percentage of granulocytic mitoses (58% to 87%) increased strikingly. These data indicate that mitotic cells of different hematopoietic cell lineages predominate in marrow at different culture times and offer a plausible explanation for the high frequency of normal karyotypes in acute myeloid leukemia after direct marrow cytogenetic evaluation.


Blood ◽  
1995 ◽  
Vol 86 (5) ◽  
pp. 1740-1748 ◽  
Author(s):  
G Klein ◽  
CA Muller ◽  
E Tillet ◽  
ML Chu ◽  
R Timpl

Collagen type VI, which forms characteristic microfibrillar structures, is assembled from three individual alpha(VI) chains that form a short triple helix and two adjacent globular domains. Expression of all three alpha (VI) collagen chains in the human bone marrow (BM) microenvironment could be detected by chain-specific antibodies in tissue sections and in the adherent stromal layer of long-term BM cultures. In functional studies, collagen type VI was shown to be a strong adhesive substrate for various hematopoietic cell lines and light-density BM mononuclear cells. The adhesive site within the molecule seems to be restricted to the triple helical domain of all three alpha (VI) chains, because individual alpha (VI) chains were not active in the attachment assays. Adhesion of the hematopoietic cell lines to collagen VI was dose-dependent and could be inhibited by heparin. Although the triple helix contains several RGD sequences, adhesion of the hematopoietic cell types to collagen VI could be blocked neither by RGD-containing peptides nor by a neutralizing antibody to the beta 1 integrin subunit. In combination with an antiadhesive substrate, the binding properties of collagen VI could be downregulated. These data suggest that this collagen type may play an important role in the adhesion of hematopoietic cells within the BM microenvironment.


1987 ◽  
Author(s):  
M C Bryckaert ◽  
A Wasteson ◽  
M Lindroth ◽  
G C Tobelem

A role for the Platelet Derived Growth Factor (PDGF) has been suggested in the abnormal proliferation of bone marrow fibroblasts occuring during myelofibrosis. To investigate this hypothesis, human bone marrow fibroblasts were isolated, and the cultures were characterized by immunofluorescent staining and electron microscopy. Electron microscopy eliminated the presence of endothelial cells by the absence of Weibel-Palade-Bodies. A positive intra and extra cellular antifibro-nectin staining was observed by immunofluorescent staining. The cultured cells didn’t show any labeling with specific antibodies for factor VIII von Willebrand factor, desmin or macrophage. Following the characterization of the bone marrow fibroblasts, using human pure 125I-PDGF, a specific binding of 125I-PDGF was demonstrated. The binding reached a plateau after 3 hours at 20°C, and after 4 hours at 4°C. Addition of unlabeled PDGF decreased this binding until 25 %.Saturation curve and scatchard analysis indicated two classes of sites with respectively 21,000 sites/aall and 37.000 sites/cell with an apparent Kd of 0.3 X 10-10 M and 0.5 X 10-9 M. Normal human serum at a concentration of 20 % induced a maximal DNA synthesis measured by-3H thymidine incorporation. When PDGF was added alone to the cultured fibroblasts at a concentration of 15 ng/ml, it induced a maximal DNA synthesis of 400 %.In the presence of 5 % of Platelet Poor Plasma (PPP), the same concentration of PDGF (15ng/ml) increased the incorporation of 3H thymidine up to 900%.In conclusion i) PDGF binds to human bone marrow fibroblasts, ii) PDGF stimulates their proliferation. These results are in favour of a role of PDGF in the proliferation of bone marrow fibroblasts associated with the development of myelofibrosis.


Blood ◽  
1990 ◽  
Vol 75 (5) ◽  
pp. 1064-1068 ◽  
Author(s):  
JM Bertho ◽  
MD Mossalayi ◽  
AH Dalloul ◽  
G Mouterde ◽  
P Debre

Abstract CD2-CD3-CD4-CD8- human bone marrow (BM) cells were previously shown to generate T-cell clones in vitro. This capacity was abolished after treatment of this population with anti-CD7 monoclonal antibody and complement. In this study, using rosetting with sheep erythrocytes, complement-dependent cytotoxicity, and specific immunoadherence method, we isolated a minor BM subset that contained more than 80% CD7+CD2-CD3- CD4-CD8- cells with small lymphoid cell morphology. They comprised most early T-cell precursors (CFU-TL) as they displayed high capacity to generate T-cell clones when cultured in limiting dilutions. CFU-TL nature of these cells was also confirmed by the sequential expression of mature T-cell specific markers on their surface after in vitro induction. This BM subset also contained 2% to 3% CFU-GM precursors. Together, these results pointed to the existence of BM CD7+CD2- precursors with high differentiation potential and showed the commitment of most of them to T-cell lineage.


2020 ◽  
Vol 12 (531) ◽  
pp. eaba9023
Author(s):  
Ioannis Zervantonakis

Human bone-marrow-on-a-chip improves drug studies of hematopoietic cell function and identifies a neutrophil maturation defect in a genetic disorder.


2020 ◽  
Vol 112 (3) ◽  
pp. 316-330
Author(s):  
Aya Fujishiro ◽  
Masaki Iwasa ◽  
Sumie Fujii ◽  
Taira Maekawa ◽  
Akira Andoh ◽  
...  

Blood ◽  
1992 ◽  
Vol 80 (4) ◽  
pp. 1074-1078 ◽  
Author(s):  
C Le Van Kim ◽  
B Cherif-Zahar ◽  
V Raynal ◽  
I Mouro ◽  
M Lopez ◽  
...  

Abstract Three Rh-related cDNAs have been isolated from a human bone marrow cDNA library and by polymerase chain reaction (PCR) amplification of human bone marrow and erythroblast mRNAs. They potentially encode a family of Rh protein isoforms that exhibit several unexpected structural properties as compared with the Rh polypeptide encoded by the cDNA clone identified previously. These modifications include several peptide deletions, the predicted alteration of Rh protein topology within the cell membrane, variations in the number and surface exposition of cysteine residues, and the generation of new C-terminal polypeptide segments caused by frameshift mutations. The four Rh mRNAs now described correspond to different splicing isoforms transcribed from the same Rh gene, and all exist in the same cell lineage (erythroid). Moreover, PCR experiments indicated that at least three of these RNA species exist in reticulocytes from donors with different commonly expressed Rh phenotypes. Although the translated proteins have not yet been characterized, these results suggest that the two genes at the RH locus may direct the synthesis of several protein species possibly corresponding to different Rh antigenic variants.


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