EC-18 (1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol) Exhibits the Immune Regulatory Role through HSC Differentiation of Bone Marrow Cells in Mice

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 4757-4757
Author(s):  
Ha-Reum Lee ◽  
Su-Hyun Shin ◽  
Nina Yoo ◽  
Sun Young Yoon ◽  
Myung-Hwan Kim ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Cell Population ◽  
Progenitor Cell ◽  
Bone Marrow Cells ◽  
Dependent Manner ◽  
Deer Antler ◽  
Myeloid Progenitor ◽  
Progenitor Cell Population ◽  
Marrow Cells

Abstract EC-18 (1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol) was originally isolated as a component of an extract from deer antler is traditionally used as an oriental medicine for hematopoiesis. A trace of MADG (monoacetyldiacylglyceride) could be detected in the deer antler, animal tissue, seed oils and bovine udder, which is identical to its natural source of MADG was chemically synthesized with glycerol, palmitic acid and linoleic acid. Previous study described that EC-18 has effect on the proliferation of hematopoietic stem cells (HSCs) (Biol. Pharm. Bull. 2004, 27(7): 1121-1125). Successively, we investigated the biological role of EC-18 in the differentiation of bone marrow cells into progenitor cell population in mice. EC-18 was administered daily for 1, 5 and 15 days to 6 week-aged C57BL/6 mice. Bone marrow cells of EC-18 administrated mice were collected from femurs and tibiae. Cell population was analyzed by FACs. As results, the total number of bone marrow cells was increased in EC-18 administered mice. To identify hematopoietic lineage cell population, we used lineage cocktail antibody kit including anti-mouse CD3, CD11b, CD45R, Gr-1 and TER-119. The lineages negative population contains HSCs which can self-renew and generate into all lineages of the hematopoietic system. The lineage negative cells significantly increased in time dependent manner in the EC-18-daily administrated mice. Cell population between myeloid progenitor (Lin- Sca1- Kit+) and common lymphoid progenitors (Lin- Sca1+ Kit+ IL-7R-α+) were analyzed using the antibody for Sca-1, c-Kit, IL-7R-α. The data showed that the population of myeloid progenitor cells was markedly increased rather than common lymphoid progenitor cells. In myeloid progenitor cell population, the cell population of megakaryocyte/erythrocyte progenitors (MEP) and granulocyte-monocyte progenitors (GMP) also elevated significantly. Taken together, EC-18 may have a potential role in the HSC differentiation and could be used as a therapeutic agent for anemia and neutropenia. Disclosures No relevant conflicts of interest to declare.

Myeloid Progenitor Cells from Gadd45a and Gadd45b-Deficient Mice Are Sensitized to Genotoxic-Stress Induced Apoptosis.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4239-4239
Author(s):  
Mamta Gupta ◽  
Shiv K. Gupta ◽  
Arthur G. Balliet ◽  
Barbara Hoffman ◽  
Dan A. Lieberman
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Bone Marrow Cells ◽  
Genotoxic Stress ◽  
Wild Type ◽  
Myeloid Progenitor ◽  
Myeloid Progenitor Cells ◽  
Induced Apoptosis ◽  
Deficient Mice ◽  
Marrow Cells

Abstract GADD45 (Growth arrest and DNA damge) regulates cell growth following exposure to diverse stimuli. It has been shown that, mice lacking the gadd45a gene exhibit genomic instability and increased carcinogenesis, but the exact role of the gadd45 family genes still remains unclear. In this study we have aimed at determining the effect of gadd45a or gadd45b deficiency on the response of bone marrow derived myeloid cells to genotoxic stress agents by using gadd45a or gadd45b null mice. We have found that myeloid progenitor cells from gadd45a or gadd45b-null mice are more sensitive to ultraviolet-radiation (UV), VP-16 or daunorubicin induced apoptosis. Introduction of wild-type gadd45 into gadd45-deficient bone marrow cells restored the wild-type apoptotic phenotype. In-vitro colony formation following stress responses has shown that bone marrow cells from gadd45a or gadd45b-deficient mice have a decreased ability to form haematopoetic colonies. Gadd45a or gadd45b-deficient bone marrow cells also displayed defective G2/M cell cycle checkpoint following exposure to either UV and V-16 but were still able to undergo G2/M arrest following exposure to daunorubicin, indicating the existence of different G2/M checkpoints in response to these anticancer agents. Taken together these findings identify gadd45a or gadd45b as anti-apoptotic gene(s), and suggests that the absence of gadd45a or gadd45b results in higher susceptibility of haematopoetic cells to UV radiation and certain anticancer drugs.

scholarly journals Abnormal responses of myeloid progenitor cells to recombinant human colony-stimulating factors in congenital neutropenia

Blood ◽  
1990 ◽  
Vol 75 (11) ◽  
pp. 2143-2149 ◽  
Author(s):  
M Kobayashi ◽  
C Yumiba ◽  
Y Kawaguchi ◽  
Y Tanaka ◽  
K Ueda ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Bone Marrow Cells ◽  
Normal Subjects ◽  
Inhibitory Effect ◽  
Congenital Neutropenia ◽  
Agar Culture ◽  
Myeloid Progenitor ◽  
Myeloid Progenitor Cells ◽  
Marrow Cells

Abstract The effects of recombinant human interleukin-3 (IL-3) and recombinant human granulocyte colony-stimulating factor (G-CSF) on the growth of myeloid progenitor cells (CFU-C) in semisolid agar culture were studied in two patients with Kostmann-type congenital neutropenia. CFU-C growth in bone marrow cells from patients was significantly reduced in response to various concentrations of either IL-3 or G-CSF alone, compared with that from normal subjects. There was no inhibitory effect of bone marrow cells from patients on normal CFU-C formation supported by IL-3 or G-CSF. However, the simultaneous stimulation with IL-3 and G- CSF induced the increase of CFU-C formation in patients with congenital neutropenia. Furthermore, CFU-C growth in both patients was supported when bone marrow cells were preincubated with IL-3 in liquid culture followed by the stimulation with G-CSF in semisolid agar culture. In contrast, that was not supported by the preincubation with G-CSF and the subsequent stimulation with IL-3. This evidence suggests that the hematopoietic progenitor cells in patients with congenital neutropenia have the potential for developing CFU-C in the combined stimulation with IL-3 and G-CSF, and that this growth may be dependent on the priming of IL-3 followed by the stimulation with G-CSF. The level of mature neutrophils in peripheral blood was not fully restored to normal levels by the daily administration of G-CSF in doses of 100 to 200 micrograms/m2 of body surface area for 20 to 25 days in both patients. These observations raise the possibility that the combination of IL-3 and G-CSF might have a potential role for the increase of neutrophil counts in patients with congenital neutropenia.

scholarly journals Abnormal responses of myeloid progenitor cells to recombinant human colony-stimulating factors in congenital neutropenia

Blood ◽  
1990 ◽  
Vol 75 (11) ◽  
pp. 2143-2149 ◽  
Author(s):  
M Kobayashi ◽  
C Yumiba ◽  
Y Kawaguchi ◽  
Y Tanaka ◽  
K Ueda ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Bone Marrow Cells ◽  
Normal Subjects ◽  
Inhibitory Effect ◽  
Congenital Neutropenia ◽  
Agar Culture ◽  
Myeloid Progenitor ◽  
Myeloid Progenitor Cells ◽  
Marrow Cells

The effects of recombinant human interleukin-3 (IL-3) and recombinant human granulocyte colony-stimulating factor (G-CSF) on the growth of myeloid progenitor cells (CFU-C) in semisolid agar culture were studied in two patients with Kostmann-type congenital neutropenia. CFU-C growth in bone marrow cells from patients was significantly reduced in response to various concentrations of either IL-3 or G-CSF alone, compared with that from normal subjects. There was no inhibitory effect of bone marrow cells from patients on normal CFU-C formation supported by IL-3 or G-CSF. However, the simultaneous stimulation with IL-3 and G- CSF induced the increase of CFU-C formation in patients with congenital neutropenia. Furthermore, CFU-C growth in both patients was supported when bone marrow cells were preincubated with IL-3 in liquid culture followed by the stimulation with G-CSF in semisolid agar culture. In contrast, that was not supported by the preincubation with G-CSF and the subsequent stimulation with IL-3. This evidence suggests that the hematopoietic progenitor cells in patients with congenital neutropenia have the potential for developing CFU-C in the combined stimulation with IL-3 and G-CSF, and that this growth may be dependent on the priming of IL-3 followed by the stimulation with G-CSF. The level of mature neutrophils in peripheral blood was not fully restored to normal levels by the daily administration of G-CSF in doses of 100 to 200 micrograms/m2 of body surface area for 20 to 25 days in both patients. These observations raise the possibility that the combination of IL-3 and G-CSF might have a potential role for the increase of neutrophil counts in patients with congenital neutropenia.

The Cell Cycle in Hematopoietic Stem Cells, Lin−/C-kit+/Sca1+ Fraction, Is Regulated by Connexin 32, Specifically Expressed in the Stem Cell Compartment

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4775-4775
Author(s):  
Yoko Hirabayashi ◽  
Byung-Il Yoon ◽  
Isao Tsuboi ◽  
Yan Huo ◽  
Yukio Kodama ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Bone Marrow ◽  
Progenitor Cells ◽  
Progenitor Cell ◽  
Bone Marrow Cells ◽  
Wild Type ◽  
Cell Compartment ◽  
Hematopoietic Stem ◽  
In The Wild ◽  
Marrow Cells

Abstract Connexin (Cx) functions in the organization of cell-cell communication via gap junctions in multicellular organisms. Gap junctions have been implicated in the homeostatic regulation of various cellular functions, including growth control, cellular differentiation, apoptosis and the synchronization of electrotonic and metabolic functions. As Cxs are essential molecules for multicellular organisms, Cxs that organize cell-cell communication within the hematopoietic progenitor cell compartment are surmised to be present in bone marrow tissue. Recently, we first found that Cx32 is only Cx molecule expressed in the bone marrow in wild-type mice by means of comparison with Cx32-knockout (KO) mice, studied by a reverse biological approach. Cx32 is specifically expressed in primitive hematopoietic stem/progenitor cells, i.e., the lineage marker-negative (Lin−)/c-kit positive (c-kit+)/stem cell antigen-1-positive (Sca1+) (=LKS) fraction, and likely playing a role of restoration of stem/progenitor cell-quiescence, thereby preventing primitive stem cells from exhaustion. In this study, we present results on cell cycle analyses with respect to the function of Cx32; one for colony-forming progenitors by the method evaluating the cycling progenitor cells using incorporation of bromodeoxyuridine (BrdUrd) followed by ultraviolet-light cytocide and the other for primitive progenitor cells using a cell sorter with bioactive AT-rich DNA-binding dye Hoechst 33342. In the colonization assay on CFU-S-13 (primitive hematopoietic progenitor cells), the incorporation of BrdUrd starts from a higher percentage with rapid increase in Cx32-KO mice, suggesting suppression of cell cycle in these primitive hematopoietic progenitor cells with Cx32-mediated cell-cycle regulation in the wild-type steady state. This suppression may be attenuated in CFU-S-9, a differentiated progenitor cell compartment. The progenitor cells assayed by in vitro colonization on CFU-GM also showed accelerated cell cycle in the Cx32-KO mice. Following the incorporation of Hoechst 33342, the lineagedepleted bone marrow cells were analyzed by flow cytometry. The population sizes of the LKS fraction obtained were 0.052% in the Cx32-KO bone marrow cells and 0.035% in the wild-type bone marrow cells (p=0.0458<0.05). The lineage-depleted bone marrow cells were analyzed their cell-cycle patterns by flow cytometry, and the G0/G1 was calculated for the LKS fractions in both, the Cx32-KO mice and wild-type mice. The percentage of G0/G1 calculated for the LKS fractions were significantly lower in the Cx32-KO mice than those in wild-type mice (60.6% vs. 87.9% for Cx32-KO vs. wild-type; p=0.001). The results suggest that Cx32 may have suppressive functions on the hematopoietic stem cell compartment, the LKS fraction, under the physiological function of Cx32. The Cx32 in the wild-type mice is, thus considered to be expressed in the primitive hematopoietic stem/progenitor cells to prevent from their exhaustion.

Toward Gene Therapy in Haemophilia A: Retrovirus-Mediated Transfer of a Factor VIII Gene into Murine Haematopoietic Progenitor Cells

1992 ◽  
Vol 67 (03) ◽  
pp. 341-345 ◽  
Author(s):  
R C Hoeben ◽  
M P W Einerhand ◽  
E Briët ◽  
H van Ormondt ◽  
D Valerio ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Bone Marrow ◽  
Factor Viii ◽  
Progenitor Cells ◽  
Progenitor Cell ◽  
Bone Marrow Cells ◽  
Coagulation Factor ◽  
Haemophilia A ◽  
Haematopoietic Progenitor Cell ◽  
Marrow Cells

SummaryTo study and evaluate the potential of the haematopoietic system as a target for gene therapy in haemophilia A, we have infected murine bone-marrow cells with a recombinant retrovirus encoding blood-coagulation factor VIII and the bacterial enzyme neomycin-phosphotransferase. After transplantation of the infected bone marrow into lethally irradiated mice, the presence of intact vector could be demonstrated in DNA isolated from individual haematopoietic progenitor-cell-derived spleen colonies. About 8% of the spleen colonies were shown to contain the intact vector. Selection for resistance to the neomycin analogue G418 prior to transplantation specifically killed the uninfected bone-marrow cells and, as a result, over 90% of the spleen colonies contained the factor VIII vector. However, expression of factor VIII in vivo, either at the RNA or at the protein level could not be demonstrated. From these data we conclude that: 1) retroviral vectors can be used to transfer factor-VIII cDNA into haematopoietic progenitor cells; 2) the vector sequences are expressed immediately after integration; and 3) transcription of the vector is repressed in the progenitor-cell-derived cells.

scholarly journals Expansion of prostate epithelial progenitor cells after inflammation of the mouse prostate

2015 ◽  
Vol 308 (12) ◽  
pp. F1421-F1430 ◽  
Author(s):  
Liang Wang ◽  
Marloes Zoetemelk ◽  
Brahmananda R. Chitteti ◽  
Timothy L. Ratliff ◽  
Jason D. Myers ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Progenitor Cells ◽  
Cell Population ◽  
Progenitor Cell ◽  
Pathological Feature ◽  
Dependent Manner ◽  
Prostate Hyperplasia ◽  
Naive Control ◽  
Progenitor Cell Population ◽  
Mouse Prostate

Prostatic inflammation is a nearly ubiquitous pathological feature observed in specimens from benign prostate hyperplasia and prostate cancer patients. The microenvironment of the inflamed prostate is highly reactive, and epithelial hyperplasia is a hallmark feature of inflamed prostates. How inflammation orchestrates epithelial proliferation as part of its repair and recovery action is not well understood. Here, we report that a novel epithelial progenitor cell population is induced to expand during inflammation. We used sphere culture assays, immunofluorescence, and flow cytometry to show that this population is increased in bacterially induced inflamed mouse prostates relative to naïve control prostates. We confirmed from previous reports that this population exclusively possesses the ability to regrow entire prostatic structures from single cell culture using renal grafts. In addition, putative progenitor cells harvested from inflamed animals have greater aggregation capacity than those isolated from naïve control prostates. Expansion of this critical cell population requires IL-1 signaling, as IL-1 receptor 1-null mice exhibit inflammation similar to wild-type inflamed animals but exhibit significantly reduced progenitor cell proliferation and hyperplasia. These data demonstrate that inflammation promotes hyperplasia in the mouse prostatic epithelium by inducing the expansion of a selected epithelial progenitor cell population in an IL-1 receptor-dependent manner. These findings may have significant impact on our understanding of how inflammation promotes proliferative diseases such as benign prostatic hyperplasia and prostate cancer, both of which depend on expansion of cells that exhibit a progenitor-like nature.

scholarly journals Detection of the target progenitor cells of granulomonopoietic enhancing activity

Blood ◽  
1990 ◽  
Vol 76 (3) ◽  
pp. 495-500
Author(s):  
SY Wang ◽  
YM Li ◽  
LY Chen ◽  
RC Wang ◽  
CK Lin ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Bone Marrow Cells ◽  
Soft Agar ◽  
Velocity Sedimentation ◽  
Gm Csf ◽  
Myeloid Progenitor ◽  
Maturation Factor ◽  
Myeloid Progenitor Cells ◽  
Marrow Cells

Macrophage-derived granulomonopoietic enhancing activity (GM-EA) is a novel mediator that amplifies colony formation of myeloid progenitor cells (CFU-GM) in conjunction with colony-stimulating factors (CSFs), and is distinct from other hematopoietic synergizing factors such as interleukin (IL)-1, IL-4, and IL-6. In the present study, we try to ascertain whether or not there is a GM-EA-specific responsive myeloid progenitor cell population. Human bone marrow cells deleted of adherent cells and T lymphocytes were separated by velocity sedimentation into three subpopulations with respective sedimentation rates (millimeters per hour) of 7.4 +/- 0.4, 6.0 +/- 0.6, and 4.7 +/- 0.3. These subpopulations corresponded to the day 7 CFU-GM, day 14 CFU-GM, and the earlier myeloid progenitor cells, pre-CFU-GM, respectively. Pre-CFU-GM failed to respond to the colony-inducing effect of GM-CSF but could be stimulated by GM-EA alone to generate small clusters (5 to 25 cells) in soft agar after 14 days of incubation. Correspondingly, suspension preculture of the fractionated bone marrow cells also showed that only the progenitor cells with low sedimentation rate (4.7 mm/h) could be activated by GM-EA to generate CFU-GM. Taken together, our results suggest that the specific target cell of GM-EA is the pre-CFU-GM, and that GM-EA acts on these cells as a growth/maturation factor, but on the day 7 and day 14 CFU-GM as a synergistic growth factor.

scholarly journals Antiapoptotic activity of Stat5 required during terminal stages of myeloid differentiation

2000 ◽  
Vol 14 (2) ◽  
pp. 232-244
Author(s):  
Matthias Kieslinger ◽  
Irina Woldman ◽  
Richard Moriggl ◽  
Johannes Hofmann ◽  
Jean-Christophe Marine ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Bone Marrow Cells ◽  
Strong Increase ◽  
Main Function ◽  
Gm Csf ◽  
Myeloid Progenitor ◽  
Myeloid Progenitor Cells ◽  
Deficient Mice ◽  
Marrow Cells

Stat5 is activated by multiple receptors of hematopoietic cytokines. To study its role during hematopoiesis, we have generated primary chicken myeloblasts expressing different dominant-negative (dn) alleles of Stat5. This caused a striking inability to generate mature cells, due to massive apoptosis during differentiation. Bcl-2 was able to rescue differentiating cells expressing dnStat5 from apoptosis, suggesting that during cytokine-dependent differentiation the main function of the protein is to ensure cell survival. Our findings with dnStat5-expressing chicken myeloblasts were confirmed with primary hematopoietic cells from Stat5a/Stat5b-deficient mice. Bone marrow cells from these animals displayed a strong increase in apoptotic cell death during GM-CSF-dependent functional maturation in vitro. The antiapoptotic protein Bcl-x was induced by GM-CSF and IL-3 in a Stat5-dependent fashion. Ectopic expression of Bcl-x rescued Stat5-deficient bone marrow cells from apoptosis, indicating that Stat5 promotes the survival of myeloid progenitor cells through its ability to induce transcription of the bcl-x gene. Finally, the recruitment of myeloid cells to inflammatory sites was found strongly impeded in Stat5-deficient mice. Taken together, our findings suggest that Stat5 may promote cytokine-dependent survival and proliferation of differentiating myeloid progenitor cells in stress or pathological situations, such as inflammation.

scholarly journals Detection of the target progenitor cells of granulomonopoietic enhancing activity

Blood ◽  
1990 ◽  
Vol 76 (3) ◽  
pp. 495-500 ◽  
Author(s):  
SY Wang ◽  
YM Li ◽  
LY Chen ◽  
RC Wang ◽  
CK Lin ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Bone Marrow Cells ◽  
Soft Agar ◽  
Velocity Sedimentation ◽  
Gm Csf ◽  
Myeloid Progenitor ◽  
Maturation Factor ◽  
Myeloid Progenitor Cells ◽  
Marrow Cells

Abstract Macrophage-derived granulomonopoietic enhancing activity (GM-EA) is a novel mediator that amplifies colony formation of myeloid progenitor cells (CFU-GM) in conjunction with colony-stimulating factors (CSFs), and is distinct from other hematopoietic synergizing factors such as interleukin (IL)-1, IL-4, and IL-6. In the present study, we try to ascertain whether or not there is a GM-EA-specific responsive myeloid progenitor cell population. Human bone marrow cells deleted of adherent cells and T lymphocytes were separated by velocity sedimentation into three subpopulations with respective sedimentation rates (millimeters per hour) of 7.4 +/- 0.4, 6.0 +/- 0.6, and 4.7 +/- 0.3. These subpopulations corresponded to the day 7 CFU-GM, day 14 CFU-GM, and the earlier myeloid progenitor cells, pre-CFU-GM, respectively. Pre-CFU-GM failed to respond to the colony-inducing effect of GM-CSF but could be stimulated by GM-EA alone to generate small clusters (5 to 25 cells) in soft agar after 14 days of incubation. Correspondingly, suspension preculture of the fractionated bone marrow cells also showed that only the progenitor cells with low sedimentation rate (4.7 mm/h) could be activated by GM-EA to generate CFU-GM. Taken together, our results suggest that the specific target cell of GM-EA is the pre-CFU-GM, and that GM-EA acts on these cells as a growth/maturation factor, but on the day 7 and day 14 CFU-GM as a synergistic growth factor.

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