Microspectrophotometric Studies of Romanowsky Stained Blood Cells. IV. Maturation of Myeloid and Erythroid Cell Lines in Bone Marrow

1984 ◽  
Vol 59 (2) ◽  
pp. 91-103 ◽  
Author(s):  
Adam Flanders ◽  
William Galbraith ◽  
Paul N. Marshall
Keyword(s):  
Bone Marrow ◽  
Cell Lines ◽  
Blood Cells ◽  
Erythroid Cell

Microrna Expression Profiling in Acute Myelogenous Leukemia.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1131-1131
Author(s):  
Fernando J. Suarez Saiz ◽  
Serban San-Marina ◽  
Mark D. Minden
Keyword(s):  
Bone Marrow ◽  
Cell Line ◽  
Cell Lines ◽  
Acute Myelogenous Leukemia ◽  
Myelogenous Leukemia ◽  
Erythroid Cell ◽  
Normal Bone Marrow ◽  
Hematopoietic Differentiation ◽  
Normal Bone ◽  
Acute Myelogenous

Abstract Acute myelogenous leukemia (AML) arises due to changes in gene expression that block or alter the normal differentiation program of hematopoietic stem cells. A variety of mutations in protein-encoding genes have been shown to contribute to the development of leukemia. Recently a new class of genes called microRNAs (miRNAs) have been identified. miRNAs are a subgroup of highly conserved, non-coding RNAs found only in eukaryotes. They do not encode proteins, and appear to have a significant effect on the proteome of a cell. Their conservation between species suggests their involvement in important biological functions, and in fact been shown to be involved in hematopoietic differentiation. While the function of most miRNAs is still unknown, it is believed that they regulate expression of target mRNAs by using the siRNA machinery either to promote degradation of the mRNA or to block its translation. To begin to understand the role of miRNAs in AML, we used Quantitative Polymerase Chain Reaction (QPCR) to measure the expression level of 20 miRNA precursors in the pro erythroid cell line K562, the pro-myelocytic cell line NB4, the myelomococytic cell line OCI/AML2, AML patients’ blasts and in normal bone marrow (NBM). The investigated miRNAs included some that are known to be specific for hematopoietic tissues or involved in hematopoietic differentiation, as well as all the miRNAs in chromosome 7, a hot spot for gene deletion in AML. Our findings indicate that miRNAs are differentially expressed in patients and cell lines when compared among themselves and against normal bone marrow. For example pre-miR-142 was expressed in NBM and K562 but was found to be elevated in OCI/AML2, NB4 and in all patient samples. Pre-miR-20 was found to be overexpressed in only a subset of patients. Other miRNAs like pre-miR-335 and pre-miR-148a were expressed in NBM and in some patients and not in the cell lines. In an effort to identify possible regulators of miRNA expression, we analyzed the upstream region of pre-miR-142 and found an LMO2 binding site. In AML, the LMO2 gene can be overexpressed relative to normal bone marrow and healthy lymphocytes. This transcription factor is involved in the regulation of genes important in the development of blood cells. To investigate if LMO2 could be involved in the regulation of miR-142 expression, we performed chromatin immunoprecipitation (ChIP) from K562 using an anti-LMO2 antibody. Only the LMO2 immunoprecipitation, and not those from the pre-immune control, were enriched in promoter DNA for pre-miR-142. This is consistent with the observation that miRNAs and coding RNAs can be regulated by the same environmental signals. Based on this observation we propose that oncogenes regulate in part the phenotype and biological behaviour of leukemia by affecting the expression of miRNAs. This further suggests that different forms of leukemia may be recognized based upon the spectrum of miRNAs they express.

scholarly journals Macrophage colony-stimulating factor (CSF-1) is expressed by spontaneously outgrown EBV-B cell lines and activated normal B lymphocytes

Blood ◽  
1989 ◽  
Vol 74 (3) ◽  
pp. 959-964
Author(s):  
G Reisbach ◽  
J Sindermann ◽  
JP Kremer ◽  
L Hultner ◽  
H Wolf ◽  
...  
Keyword(s):  
Bone Marrow ◽  
B Cell ◽  
Cell Line ◽  
B Lymphocytes ◽  
Cell Lines ◽  
Blood Cells ◽  
Colony Stimulating Factor ◽  
Normal Human ◽  
Macrophage Colony ◽  
Stimulating Factor

Human B lymphocytes activated by mitogens or infected by Epstein Barr virus (EBV) have previously been shown to release colony-stimulating activity (CSA) supporting the growth of normal human bone marrow progenitors. We established five different human EBV-B cell lines spontaneously outgrown from nonmalignant peripheral blood cells and long-term bone marrow cultures. CSA derived from all of these lines induces the growth of murine macrophage colonies, whereas virtually no human bone marrow cell progenitors were stimulated. As observed in the tumor cell line MIA PaCa-2, a 4.3-kilobase (kb) transcript was detected in all cases using a human colony-stimulating factor (CSF)-1 probe. Expression of this transcript can be further stimulated within three hours upon addition of phorbol myristate acetate (PMA). The highly purified native protein exerting macrophage colony-stimulating activity (M-CSA) exhibits a molecular size of approximately 75 to 97 Kd in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The identity of EBV-B cell derived M-CSA with human urinary CSF-1 was confirmed by a complete neutralization of macrophage CSA by an antihuman urinary CSF-1 antiserum. Normal human B lymphocytes purified from tonsils or from mononuclear blood cells also express CSF-1 upon stimulation with Staphylococcus aureus Cowan I. No CSF-1 expression, however, could be detected in normal resting B lymphocytes or in the Burkitt lymphoma cell line RAJI.

scholarly journals Stress erythropoiesis in atherogenic mice

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Ángela Sánchez ◽  
Marta C. Orizaola ◽  
Diego Rodríguez-Muñoz ◽  
Ana Aranda ◽  
Antonio Castrillo ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Apolipoprotein E ◽  
Blood Cells ◽  
High Fat Diet ◽  
Cell Lineage ◽  
Erythroid Cell ◽  
High Fat ◽  
Erythroid Progenitors ◽  
Stress Erythropoiesis ◽  
Colony Forming Unit

Abstract Bone marrow erythropoiesis is mainly homeostatic and a demand of oxygen in tissues activates stress erythropoiesis in the spleen. Here, we show an increase in the number of circulating erythrocytes in apolipoprotein E−/− mice fed a Western high-fat diet, with similar number of circulating leukocytes and CD41+ events (platelets). Atherogenic conditions increase spleen erythropoiesis with no variations of this cell lineage in the bone marrow. Spleens from atherogenic mice show augmented number of late-stage erythroblasts and biased differentiation of progenitor cells towards the erythroid cell lineage, with an increase of CD71+CD41CD34−CD117+Sca1−Lin− cells (erythroid-primed megakaryocyte-erythroid progenitors), which is consistent with the way in which atherogenesis modifies the expression of pro-erythroid and pro-megakaryocytic genes in megakaryocyte-erythroid progenitors. These data explain the transiently improved response to an acute severe hemolytic anemia insult found in atherogenic mice in comparison to control mice, as well as the higher burst-forming unit-erythroid and colony forming unit-erythroid capacity of splenocytes from atherogenic mice. In conclusion, our work demonstrates that, along with the well stablished enhancement of monocytosis during atherogenesis, stress erythropoiesis in apolipoprotein E−/− mice fed a Western high fat diet results in increased numbers of circulating red blood cells.

The Expression and Regulation of ADAMTS13 in Hematopoietic System.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4438-4438
Author(s):  
Fang Liu ◽  
Xia Bai ◽  
Lan Dai Master ◽  
Ningzheng Dong ◽  
Changgeng Ruan
Keyword(s):  
Bone Marrow ◽  
Cell Lines ◽  
Blood Cells ◽  
Major Part ◽  
Positive Control ◽  
Human Platelets ◽  
Blast Cell ◽  
Rt Pcr ◽  
Hematopoietic System ◽  
Variable Expression

Abstract Background and Objective In 2001, VWF-cleaving protease was identified as ADAMTS13, a member of the ADAMTS family of metalloprotease. ADAMTS13 is now found as the mainly cleaving enzyme of VWF in plasma. It is synthesized predominantly in the liver, although variable expression has been observed in other issues including bone marrow. Although ADAMTS13 was detected only in platelet, not other blood cells, it is unsure what kind of situations would be when we come to immature hematopoietic cells. So we investigated ADAMTS13 in cell lines such as M07e, MEG01, HEL, k562, SHI-1, NB4, MR2 etc., to see if there is ADAMTS13 produced by blast cell of megakaryocyte and granulocyte and so on. Methods We determined ADAMTS13 cDNA by RT-PCR and ADAMTS13 expression by flow cytometry and western blotting with some mono-clone antibodies against ADAMTS13. During all those test, we use human platelets as positive control. Results ADAMTS13 was detectable in all those cell lines. It was highly expressed in M07e and MR2, and a bit lower in MEG01 and HEL, and much lower in k562, SHI-1 and NB4. While to our surprise, ADAMTS13 was up-regulated when NB4 was treated by 1μmol/L ATRA for 72 hours. When CD11b was rising from 21% up to 95% in NB4 after treatment, expression of ADAMTS13 followed by from 24% to 80%. Conclusions Now we know what kinds of cells may express ADAMTS13 in bone marrow, the major part of it is localized in megakaryocyte. ADAMTS13 can highly expressed in NB4 after treatment with ARTA.

scholarly journals Plasticity of Cells andEx VivoProduction of Red Blood Cells

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Takashi Hiroyama ◽  
Kenichi Miharada ◽  
Ryo Kurita ◽  
Yukio Nakamura
Keyword(s):  
Red Blood Cells ◽  
Cell Lines ◽  
Blood Cells ◽  
Progenitor Cell ◽  
Ex Vivo ◽  
Es Cells ◽  
Erythroid Cell ◽  
Erythroid Progenitor ◽  
Hematopoietic Stem ◽  
Erythroid Progenitor Cell

The supply of transfusable red blood cells (RBCs) is not sufficient in many countries. If transfusable RBCs could be produced abundantly from certain resources, it would be very useful. Our group has developed a method to produce enucleated RBCs efficiently from hematopoietic stem/progenitor cells present in umbilical cord blood. More recently, it was reported that enucleated RBCs could be abundantly produced from human embryonic stem (ES) cells. The common obstacle for application of these methods is that they require very high cost to produce sufficient number of RBCs that are applicable in the clinic. If erythroid cell lines (immortalized cell lines) able to produce transfusable RBCsex vivowere established, they would be valuable resources. Our group developed a robust method to obtain immortalized erythroid cell lines able to produce mature RBCs. To the best of our knowledge, this was the first paper to show the feasibility of establishing immortalized erythroid progenitor cell lines able to produce enucleated RBCsex vivo. This result strongly suggests that immortalized human erythroid progenitor cell lines able to produce mature RBCsex vivocan also be established.

scholarly journals Macrophage colony-stimulating factor (CSF-1) is expressed by spontaneously outgrown EBV-B cell lines and activated normal B lymphocytes

Blood ◽  
1989 ◽  
Vol 74 (3) ◽  
pp. 959-964 ◽  
Author(s):  
G Reisbach ◽  
J Sindermann ◽  
JP Kremer ◽  
L Hultner ◽  
H Wolf ◽  
...  
Keyword(s):  
Bone Marrow ◽  
B Cell ◽  
Cell Line ◽  
B Lymphocytes ◽  
Cell Lines ◽  
Blood Cells ◽  
Colony Stimulating Factor ◽  
Normal Human ◽  
Macrophage Colony ◽  
Stimulating Factor

Abstract Human B lymphocytes activated by mitogens or infected by Epstein Barr virus (EBV) have previously been shown to release colony-stimulating activity (CSA) supporting the growth of normal human bone marrow progenitors. We established five different human EBV-B cell lines spontaneously outgrown from nonmalignant peripheral blood cells and long-term bone marrow cultures. CSA derived from all of these lines induces the growth of murine macrophage colonies, whereas virtually no human bone marrow cell progenitors were stimulated. As observed in the tumor cell line MIA PaCa-2, a 4.3-kilobase (kb) transcript was detected in all cases using a human colony-stimulating factor (CSF)-1 probe. Expression of this transcript can be further stimulated within three hours upon addition of phorbol myristate acetate (PMA). The highly purified native protein exerting macrophage colony-stimulating activity (M-CSA) exhibits a molecular size of approximately 75 to 97 Kd in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The identity of EBV-B cell derived M-CSA with human urinary CSF-1 was confirmed by a complete neutralization of macrophage CSA by an antihuman urinary CSF-1 antiserum. Normal human B lymphocytes purified from tonsils or from mononuclear blood cells also express CSF-1 upon stimulation with Staphylococcus aureus Cowan I. No CSF-1 expression, however, could be detected in normal resting B lymphocytes or in the Burkitt lymphoma cell line RAJI.

scholarly journals Targeted expression of a conditional oncogene in hematopoietic cells of transgenic mice.

1993 ◽  
Vol 123 (6) ◽  
pp. 1545-1553 ◽  
Author(s):  
K Ravid ◽  
Y C Li ◽  
H B Rayburn ◽  
R D Rosenberg
Keyword(s):  
Bone Marrow ◽  
Transgenic Mice ◽  
Cell Lines ◽  
T Antigen ◽  
Erythroid Cell ◽  
Specific Gene ◽  
In Vitro Cultivation ◽  
Permissive Temperature ◽  
Cell Marker ◽  
Platelet Factor

We have produced two lines of transgenic mice in which the expression of temperature-sensitive SV-40 large T antigen is targeted to bone marrow megakaryocytes via the platelet factor 4 (PF4) tissue-specific promoter. The progeny of these transgenic mice were observed for about 3 mo, and no malignancies were detected over this period of time. The offspring of these transgenic mice, 6- to 12-wk of age, served as a source of bone marrow cells, which upon in vitro cultivation at the permissive temperature yielded immortalized cell lines (MegT). At the permissive temperature, MegT cells exhibit the characteristics of early 2N and 4N megakaryocytes which include the presence of specific gene products such as PF4, glycoprotein IIb, acetylcholinesterase, and CD45 as well as the absence of molecular markers of other cell lineages such as the macrophage marker Mac-1, the T helper cell marker CD4, the mast cell marker IgE, the T cell marker CD2 or the erythroid cell marker alpha-globin. The inactivation of the oncogene by a shift of temperature from 34 degrees to 39.5 degrees C produces a reduction in the frequency of the 2N cells, in conjunction with the appearance of 8N and 16N cells, consisting of 27 and 3% of total cells, respectively. Thus, we have generated hematopoietic cell lines that are trapped in the early stages of megakaryocyte commitment, but able to undergo part of the normal program of terminal differentiation.

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