Fetal murine hemopoiesis following in utero low-dose irradiation

Development ◽  
1981 ◽  
Vol 62 (1) ◽  
pp. 37-46
Author(s):  
Sheila R. Weinberg ◽  
Emmeline G. McCarthy ◽  
Thomas J. MacVittie
Keyword(s):  
Low Dose ◽  
Fetal Liver ◽  
Soft Agar ◽  
In Utero ◽  
In Vitro Assays ◽  
Plasma Clot ◽  
Erythroid Progenitor ◽  
Colony Forming Cell ◽  
Macrophage Colony

The influence of in utero low-dose ionizing radiation exposure on murine hemopoietic embryogenesis was investigated. In vitro assays such as micro plasma-clot cultures and double-layer soft agar cultures served as sensitive biodosimeters to determine erythropoietic and granulopoietic injuries. Day-10·5, HA/ICR, pregnant mice were irradiated with 0, 50, 100, 150,200, or 300 rads, and day-14·5 fetal livers were studied for colony-forming unit-erythroid (CFU-E), buist-forming unit-erythroid (BFU-E), granulocyte-macrophage colony-forming cell (GM-CFC), and macrophage-colony-forming cell (M-CFC) activity. Fetuses subjected to doses of 200 rads or higher on day 10·5 of gestation responded with a decrease in day-14·5 liver cellularity, reflecting injury to the developing organ and its inability to recover to the nonirradiated values. Difference in response between erythropoietin(EPO)-dependent and EPO-independent CFU-E strongly suggests existence of two populations of erythroid progenitor cells with different radiosensitivities. A dose of 200 rads markedly reduced CFUE recovery, and a dose of 100 rads was sufficient to reduce BFU-E recovery to almost 10% of 0-rad values. Nonirradiated day-14·5 fetal liver had more GM-CFC compared to any of the irradiated fetuses, and a dramatically reduced M-CFC recovery occurred with each increase in dose following 150 rads. Our results showed that (1) fetal liver granulopoiesis is more sensitive to radiation injury compared to erythropoiesis, and (2) fetal liver has a greater potential for erythropoiesis recovery.

The relationship between erythropoietin-dependent cellular differentiation and colony-forming ability in prenatal haemopoietic tissues

Development ◽  
1975 ◽  
Vol 34 (3) ◽  
pp. 575-588
Author(s):  
R. J. Cole ◽  
T. Regan ◽  
S. L. White ◽  
E. M. Cheek
Keyword(s):  
Cellular Differentiation ◽  
Cell Numbers ◽  
In Vitro Sensitivity ◽  
Foetal Liver ◽  
Rapid Production ◽  
Colony Forming Cell ◽  
Macrophage Colony ◽  
The Relationship

Levels of haem synthesis achieved by foetal liver erythroblasts responding to erythropoietin in vitro are similar in dissociated cell cultures and in cultures of organized tissues. Erythroid colony-forming cells reach maximum numbers on the sixteenth day of gestation. Their presence in foetal liver is associated with the period of most rapid production of erythrocytes, and with in vitro sensitivity to erythropoietin measured as enhanced haem synthesis. It is concluded that at least a proportion of erythroid colony-forming cells in the foetal liver are dependent on erythropoietin in situ and that these cells are separated from the earliest recognizable pro-erythroblast by 1–2 cell divisions. Populations of granulocyte-macrophage colony-forming cells change independently of erythroid colony-forming cell numbers.

scholarly journals A critical role of VLA-4 in erythropoiesis in vivo

Blood ◽  
1996 ◽  
Vol 87 (6) ◽  
pp. 2513-2517 ◽  
Author(s):  
K Hamamura ◽  
H Matsuda ◽  
Y Takeuchi ◽  
S Habu ◽  
H Yagita ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Fetal Liver ◽  
Critical Role ◽  
In Utero ◽  
In Vitro Studies ◽  
Specific Interactions ◽  
Erythroid Progenitors

Hematopoiesis requires specific interactions with the microenvironments, and VLA-4 has been implicated in these interactions based on in vitro studies. To study the role of VLA-4 in hematopoiesis in vivo, we performed in utero treatment of mice with an anti-VLA-4 monoclonal antibody. Although all hematopoietic cells in fetal liver expressed VLA-4, the treatment specifically induced anemia. It had no effect on the development of nonerythroid lineage cells, including lymphoids and myeloids. In the treated liver almost no erythroblast was detected, whereas the erythroid progenitors, which give rise to erythroid colonies in vitro, were present. These results indicate that VLA-4 plays a critical role in erythropoiesis, while it is not critical in lymphopoiesis in vivo.

scholarly journals A bleeding disorder due to deficiency of alpha 2-antiplasmin

Blood ◽  
1982 ◽  
Vol 59 (6) ◽  
pp. 1246-1251 ◽  
Author(s):  
LA Miles ◽  
EF Plow ◽  
KJ Donnelly ◽  
C Hougie ◽  
JH Griffin
Keyword(s):  
System Analysis ◽  
Bleeding Disorder ◽  
Fibrinolytic System ◽  
In Vitro Assays ◽  
Clot Lysis ◽  
Plasma Clot ◽  
Normal Limits ◽  
Normal Plasma ◽  
Bleeding Episodes

Abstract A deficiency of alpha 2-antiplasmin has been identified in a female patient with severe and frequent bleeding episodes. Routine coagulation and platelet assays of the patient's plasma were within normal limits. However, abnormally rapid whole blood or dilute plasma clot lysis times and an abnormal FXIII test in which clots were lysed in the presence of urea or saline suggested an abnormal fibrinolytic system. Analysis of alpha 2-antiplasmin levels by radioimmunoassay revealed less than 1.0 microgram/ml alpha 2-antiplasmin. Functional assays indicated an alpha 2-antiplasmin level less than or equal to 10% of normal. Addition of purified alpha 2-antiplasmin to the patient's plasma restored its ability to inhibit plasmin in in vitro assays, and mixtures of patient plasma with normal plasma did not interfere with the antiplasmin activity of the normal plasma. Whereas normal platelets contain 68 ng alpha 2-antiplasmin/10(9) platelets, platelets from the patient contained 30% of the normal level of antigen. Analysis of alpha 2- antiplasmin functional and antigenic levels in the plasma of both parents and four siblings of the propositus provided evidence consistent with an autosomal mechanism of inheritance of alpha 2- antiplasmin deficiency. One sibling appeared to be homozygous and three siblings and the parents were heterozygous for the deficiency. Two heterozygotes had positive bleeding histories. The association of a bleeding disorder with a deficiency of alpha 2-antiplasmin emphasizes that lack of regulation of the fibrinolytic system can result in a hemostatic dysfunction.

scholarly journals Chronic Microcystin-LR Exposure Induces Hepatocarcinogenesis via Increased Gankyrin in Vitro and in Vivo

2018 ◽  
Vol 49 (4) ◽  
pp. 1420-1430 ◽  
Author(s):  
Lixiong He ◽  
Yujing Huang ◽  
Qiaonan Guo ◽  
Hui Zeng ◽  
Chuanfen Zheng ◽  
...  
Keyword(s):  
Low Dose ◽  
Soft Agar ◽  
Tumor Formation ◽  
Sprague Dawley ◽  
Sprague Dawley Rats ◽  
L02 Cells ◽  
Insight Into

Background/Aims: Our recent study indicated that the serum microcystin-LR (MC-LR) level is positively linked to the risk of human hepatocellular carcinoma (HCC). Gankyrin is over-expressed in cancers and mediates oncogenesis; however, whether MC-LR induces tumor formation and the role of gankyrin in this process is unclear. Methods: We induced malignant transformation of L02 liver cells via 35 passages with exposure to 1, 10, or 100 nM MC-LR. Wound healing, plate and soft agar colony counts, and nude mice tumor formation were used to evaluate the tumorigenic phenotype of MC-LR-treated cells. Silencing gankyrin was used to confirm its function. We established a 35-week MC-LR exposure rat model by twice weekly intraperitoneal injection with 10 μg/kg body weight. In addition, 96 HCC patients were tested for tumor tissue gankyrin expression and serum MC-LR levels. Results: Chronic low-dose MC-LR exposure increased proliferation, mobility, clone and tumor formation abilities of L02 cells as a result of gankyrin activation, while silencing gankyrin inhibited the carcinogenic phenotype of MC-LR-treated cells. MC-LR also induced neoplastic liver lesions in Sprague-Dawley rats due to up-regulated gankyrin. Furthermore, a trend of increased gankyrin was observed in humans exposed to MC-LR. Conclusion: These results suggest that MC-LR induces hepatocarcinogenesis in vitro and in vivo by increasing gankyrin levels, providing new insight into MC-LR carcinogenicity studies.

Influence of albuterol on erythropoietin production and erythroid progenitor cell activation

1979 ◽  
Vol 236 (3) ◽  
pp. H422-H426 ◽  
Author(s):  
F. Przala ◽  
D. M. Gross ◽  
B. Beckman ◽  
J. W. Fisher
Keyword(s):  
Bone Marrow ◽  
Progenitor Cell ◽  
Cell Activation ◽  
Plasma Clot ◽  
Erythroid Progenitor ◽  
Erythroid Progenitor Cell ◽  
Adrenergic Blocker ◽  
Rabbit Bone

The effect of albuterol, a potent beta2-adrenergic agonist, on kidney production of erythropoietin (Ep) was studied. Its effects on erythroid colony (CFU-E) formation in vitro in rabbit bone marrow cultures were also assessed. Albuterol produced a significant increase in plasma Ep levels in conscious rabbits following 7 h intravenous infusion (50 (microgram/kg)/min). This effect was blocked by pretreatment of the rabbits with butoxamine (5 mg/kg ip), a potent beta2-adrenergic blocker. Albuterol in doses of 10(-10) to 10(-8) M in combination with Ep was also found to produce a significant increase in the numbers of CFU-E in the plasma clot culture system of rabbit bone marrow. This effect was blocked completely by DL-propranolol (10(-8) M) and by butoxamine (10(-8) M). The data presented suggest that albuterol, a potent activator of beta2-adrenergic receptors, increases kidney production of Ep in vivo and also produces a direct effect in combination with Ep on the proliferation of the erythroid progenitor cell compartment.

scholarly journals Low dose amiodarone reduces tumor growth and angiogenesis

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Eliana Steinberg ◽  
Arnon Fluksman ◽  
Chalom Zemmour ◽  
Katerina Tischenko ◽  
Adi Karsch-Bluman ◽  
...  
Keyword(s):  
Side Effects ◽  
Cancer Cells ◽  
Low Dose ◽  
Tumor Model ◽  
In Vitro Assays ◽  
Potential Candidate ◽  
Key Factor ◽  
Angiogenic Effect

Abstract Amiodarone is an anti-arrhythmic drug that was approved by the US Food and Drug Administration (FDA) in 1985. Pre-clinical studies suggest that Amiodarone induces cytotoxicity in several types of cancer cells, thus making it a potential candidate for use as an anti-cancer treatment. However, it is also known to cause a variety of severe side effects. We hypothesized that in addition to the cytotoxic effects observed in cancer cells Amiodarone also has an indirect effect on angiogensis, a key factor in the tumor microenvironment. In this study, we examined Amiodarone's effects on a murine tumor model comprised of U-87 MG glioblastoma multiforme (GBM) cells, known to form highly vascularized tumors. We performed several in vitro assays using tumor and endothelial cells, along with in vivo assays utilizing three murine models. Low dose Amiodarone markedly reduced the size of GBM xenograft tumors and displayed a strong anti-angiogenic effect, suggesting dual cancer fighting properties. Our findings lay the ground for further research of Amiodarone as a possible clinical agent that, used in safe doses, maintains its dual properties while averting the drug’s harmful side effects.

scholarly journals Pregnancy-Secreted Acid Phosphatase, Uteroferrin, Enhances Fetal Erythropoiesis

Endocrinology ◽  
2014 ◽  
Vol 155 (11) ◽  
pp. 4521-4530 ◽  
Author(s):  
Wei Ying ◽  
Haiqing Wang ◽  
Fuller W. Bazer ◽  
Beiyan Zhou
Keyword(s):  
Acid Phosphatase ◽  
Progenitor Cells ◽  
Fetal Liver ◽  
Hematopoietic Cells ◽  
Erythroid Progenitor ◽  
Colony Forming Unit ◽  
Erythroid Progenitor Cells ◽  
Uterine Glands ◽  
Fetal Erythropoiesis

Abstract Uteroferrin (UF) is a progesterone-induced acid phosphatase produced by uterine glandular epithelia in mammals during pregnancy and targeted to sites of hematopoiesis throughout pregnancy. The expression pattern of UF is coordinated with early fetal hematopoietic development in the yolk sac and then liver, spleen, and bone to prevent anemia in fetuses. Our previous studies suggested that UF exerts stimulatory impacts on hematopoietic progenitor cells. However, the precise role and thereby the mechanism of action of UF on hematopoiesis have not been investigated previously. Here, we report that UF is a potent regulator that can greatly enhance fetal erythropoiesis. Using primary fetal liver hematopoietic cells, we observed a synergistic stimulatory effect of UF with erythropoietin and other growth factors on both burst-forming unit-erythroid and colony-forming unit-erythroid formation. Further, we demonstrated that UF enhanced erythropoiesis at terminal stages using an in vitro culture system. Surveying genes that are crucial for erythrocyte formation at various stages revealed that UF, along with erythropoietin, up-regulated transcription factors required for terminal erythrocyte differentiation and genes required for synthesis of hemoglobin. Collectively, our results demonstrate that UF is a cytokine secreted by uterine glands in response to progesterone that promotes fetal erythropoiesis at various stages of pregnancy, including burst-forming unit-erythroid and colony-forming unit-erythroid progenitor cells and terminal stages of differentiation of hematopoietic cells in the erythroid lineage.

scholarly journals Clarifying the role of Stat5 in lymphoid development and Abelson-induced transformation

Blood ◽  
2006 ◽  
Vol 107 (12) ◽  
pp. 4898-4906 ◽  
Author(s):  
Andrea Hoelbl ◽  
Boris Kovacic ◽  
Marc A. Kerenyi ◽  
Olivia Simma ◽  
Wolfgang Warsch ◽  
...  
Keyword(s):  
B Cell ◽  
Fetal Liver ◽  
Transgenic Animals ◽  
Cell Receptor ◽  
Lymphoid Cells ◽  
In Vitro Assays ◽  
Cell Stage ◽  
Lymphoid Development ◽  
B Lymphoid

AbstractThe Stat5 transcription factors Stat5a and Stat5b have been implicated in lymphoid development and transformation. Most studies have employed Stat5a/b-deficient mice where gene targeting disrupted the first protein-coding exon, resulting in the expression of N-terminally truncated forms of Stat5a/b (Stat5a/bΔN/ΔN mice). We have now reanalyzed lymphoid development in Stat5a/bnull/null mice having a complete deletion of the Stat5a/b gene locus. The few surviving Stat5a/bnull/null mice lacked CD8+ T lymphocytes. A massive reduction of CD8+ T cells was also found in Stat5a/bfl/fllck-cre transgenic animals. While γδ T-cell receptor–positive (γδTCR+) cells were expressed at normal levels in Stat5a/bΔN/ΔN mice, they were completely absent in Stat5a/bnull/null animals. Moreover, B-cell maturation was abrogated at the pre–pro-B-cell stage in Stat5a/bnull/null mice, whereas Stat5a/bΔN/ΔN B-lymphoid cells developed to the early pro-B-cell stage. In vitro assays using fetal liver-cell cultures confirmed this observation. Most strikingly, Stat5a/bnull/null cells were resistant to transformation and leukemia development induced by Abelson oncogenes, whereas Stat5a/bΔN/ΔN-derived cells readily transformed. These findings show distinct lymphoid defects for Stat5a/bΔN/ΔN and Stat5a/bnull/null mice and define a novel functional role for the N-termini of Stat5a/b in B-lymphoid transformation.

UCP2 Modulates Cell Proliferation through the MAPK/ERK Pathway during Erythropoiesis and Has No Effect on Heme Biosynthesis

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 5372-5372
Author(s):  
Alvaro A Elorza ◽  
Brigham B Hyde ◽  
Hanna Mikkola ◽  
Sheila Collins ◽  
Orian S Shirihai
Keyword(s):  
Cell Proliferation ◽  
Progenitor Cells ◽  
Fetal Liver ◽  
Mitochondrial Protein ◽  
Erythroid Progenitor ◽  
Heme Synthesis ◽  
Erythroid Progenitor Cells ◽  
Deficient Mice

Abstract UCP2, an inner membrane mitochondrial protein, has been implicated in bioenergetics and Reactive Oxygen Species (ROS) modulation. UCP2 has been previously hypothesized to function as a facilitator of heme synthesis and iron metabolism by reducing ROS production. While UCP2 has been found to be induced by GATA1 during erythroid differentiation its role in erythropoiesis in vivo or in vitro has not been reported thus far. Here we report on the study of UCP2 role in erythropoiesis and the hematologic phenotype of UCP2 deficient mouse. In vivo we found that UCP2 protein peaks at early stages of erythroid maturation when cells are not fully committed in heme synthesis and then becomes undetectable at the reticulocyte stage. Iron incorporation into heme was unaltered in erythroid cells from UCP2 deficient mice. While heme synthesis was not influenced by UCP2 deficiency, mice lacking UCP2 had a delayed recovery from chemically induced hemolytic anemia. Analysis of the erythroid lineage from bone marrow and fetal liver revealed that in the UCP2 deficient mice the R3 (CD71high/Ter119high) population was reduced by 24%. The count of BFU-E and CFU-E colonies, scored in an erythroid colony assay, was unaffected, indicating an equivalent number of early erythroid progenitor cells in both UCP2 deficient and control cells. Ex-vivo differentiation assay revealed that UCP2 deficient c-kit+ progenitor cells expansion was overall reduced by 14% with population analysis determining that the main effect is at the R3 stage. No increased rate of apoptosis was found indicating that expansion rather than cell death is being compromised. Reduced expansion of c-kit+ cells was accompanied by 30% reduction in the phosphorylated form of ERK, a ROS dependent cytosolic regulator of cell proliferation. Analysis of ROS in UCP2 null erythroid progenitors revealed altered distribution of ROS resulting in 14% decrease in cytosolic and 32% increase in mitochondrial ROS. Restoration of the cytosolic oxidative state of erythroid progenitor cells by the pro-oxidant Paraquat reversed the effect of UCP2 deficiency on cell proliferation in in vitro differentiation assays. Together, these results indicate that UCP2 is a regulator of erythropoiesis and suggests that inhibition of UCP2 function may contribute to the development of anemia.

Growth suppression of human pancreatic cells by a novel PKCβ inhibitor, enzastaurin

2007 ◽  
Vol 25 (18_suppl) ◽  
pp. 14141-14141
Author(s):  
H. Cheng ◽  
J. G. Yan ◽  
Y. Jiang
Keyword(s):  
Pancreatic Cancer ◽  
Cell Proliferation ◽  
Mtt Assay ◽  
Low Dose ◽  
Soft Agar ◽  
Human Pancreatic Cancer ◽  
Pancreatic Cancer Cells ◽  
Pancreatic Cells ◽  
Transforming Activity

14141 Background: Human pancreatic cancer is a devastating disease. Standard therapy, gemcitabine, is not able to alter the natural history of the disease. Targeting intracellular signaling molecules has proven to be a powerful approach in cancer therapy. Protein kinase C (PKC) family has been implicated in many human cancers including pancreatic cancer. Methods: In this pre-clinical study, we evaluated the effect of a novel small molecule inhibitor, enzastaurin, for PKCβ on human pancreatic cancer cells. We utilized MTT assay to assess cell proliferation and viability. To evaluate cell transforming activity, soft agar colony forming assay was used. Results: We showed that with addition of 20μM enzastaurin into the culture, the proliferative growth of two pancreatic cancer cell lines, Panc1 and L3.6p1, was significantly attenuated, as measured by the MTT assay. In addition, the combination of enzastaurin and a low dose of gemcitabine (25 nM) resulted in a synergistic cytotoxic effect in L3.6p1 cells. Further, the in vitro tumorogenicity assay demonstrated that the L3.6p1 cells treated with enzastaurin (20μM) formed much smaller colonies than the control parental cells in 0.4% soft-agar. Consistent with the in vitro cell proliferation results, the combination of enzastaurin and the low concentration of gemcitabine (25nM) further reduced the numbers and sizes of colonies in the soft-agar. Conclusions: Collectively, our data indicate that enzastaurin can effectively suppress the proliferation and in vitro transforming activity of the pancreatic cells, and more effectively when applied in combination with the low dose of gemcitabine. No significant financial relationships to disclose.

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