STIMULATION OF BONE MARROW CELLS AND BONE FORMATION BY NACRE. IN VIVO AND IN VITRO STUDIES

Liquid cultures of bone marrow cells enriched for megakaryocytes were assayed for incorporation of 3H-thymidine (3H-TdR) into acid-precipitable cell digests to determine the effect of thrombopoietin on DNA synthesis. As previously described, thrombopoietin was prepared by ammonium sulfate fractionation of pooled plasma obtained from thrombocytopenic rabbits. A control fraction was prepared from normal rabbit plasma. The thrombopoietic activity of these fractions was determined in vivo with normal rabbits as assay animals and the rate of incorporation of 75Se-selenomethionine into newly formed platelets as an index of thrombopoietic activity of the infused material. Guinea pig megakaryocytes were purified using bovine serum albumin gradients. Bone marrow cultures containing 1.5-3.0x104 cells and 31%-71% megakaryocytes were incubated 18 h in modified Dulbecco’s MEM containing 10% of the concentrated plasma fractions from either thrombocytopenic or normal rabbits. In other control cultures, 0.9% NaCl was substituted for the plasma fractions. 3H-TdR incorporation was measured after cells were incubated for 3 h with 1 μCi/ml. The protein fraction containing thrombopoietin-stimulating activity caused a 25%-31% increase in 3H-TdR incorporation over that in cultures which were incubated with the similar fraction from normal plasma and a 29% increase over the activity in control cultures to which 0.9% NaCl had been added. These data suggest that thrombopoietin stimulates DNA synthesis in megakaryocytes and that this tecnique may be useful in assaying thrombopoietin in vitro.

Download Full-text

In vitro stimulation of primate hemoglobin synthesis by L-thyroxine

Blood ◽

10.1182/blood.v49.3.407.407 ◽

1977 ◽

Vol 49 (3) ◽

pp. 407-413 ◽

Cited By ~ 4

Author(s):

JE Fuhr ◽

N Gengozian ◽

M Overton

Keyword(s):

Bone Marrow ◽

Bone Marrow Cells ◽

Leucine Incorporation ◽

Hemoglobin Synthesis ◽

Globin Chains ◽

Stimulation Of ◽

Marrow Cells

Abstract Bone marrow cells from adult and abortus primates (marmosets) were incubated in vitro to determine their responsiveness to L-thyroxine. 3H- leucine incorporation into purified globin chains was the parameter assayed to determine responsiveness. Bone marrow from spontaneously aborted animals consistently was stimulated by the presence of physiologic levels of L-thyroxine. Bone marrow cells from adult animals were unaffected by the hormone.

Download Full-text

IN VITRO ASSAY OF CYTOGENETIC DAMAGE INDUCED IN BONE MARROW CELLS IN VIVO BY CHEMICAL CARCINOGENS

Japanese Journal of Medical Science and Biology ◽

10.7883/yoken1952.38.207 ◽

1985 ◽

Vol 38 (5-6) ◽

pp. 207-216 ◽

Cited By ~ 3

Author(s):

Esmail Kodhom SHUBBER ◽

David KRAM ◽

Jerry WILLIAMS

Keyword(s):

Bone Marrow ◽

Bone Marrow Cells ◽

In Vitro Assay ◽

Chemical Carcinogens ◽

Vitro Assay ◽

Cytogenetic Damage ◽

Marrow Cells

Download Full-text

Coencapsulation of hepatocytes and bone marrow cells: In vitro and in vivo studies

Biotechnology Annual Review ◽

10.1016/s1387-2656(06)12005-0 ◽

2006 ◽

pp. 137-151 ◽

Cited By ~ 11

Author(s):

Zun Chang Liu ◽

Thomas Ming Swi Chang

Keyword(s):

Bone Marrow ◽

Bone Marrow Cells ◽

In Vivo Studies ◽

Marrow Cells

Download Full-text

Increase in the Number of Bone Marrow Osteoclast Precursors at Different Skeletal Sites, Particularly in Long Bone and Jaw Marrow in Mice Lacking IL-1RA

International Journal of Molecular Sciences ◽

10.3390/ijms21113774 ◽

2020 ◽

Vol 21 (11) ◽

pp. 3774

Author(s):

Giuliana Ascone ◽

Yixuan Cao ◽

Ineke D.C. Jansen ◽

Irene Di Ceglie ◽

Martijn H.J. van den Bosch ◽

...

Keyword(s):

Bone Marrow ◽

Bone Marrow Cells ◽

Interleukin 1 ◽

Bone Destruction ◽

Mineral Dissolution ◽

Long Bone ◽

Osteoclast Precursors ◽

Marrow Cells

Recently, it was shown that interleukin-1β (IL-1β) has diverse stimulatory effects on different murine long bone marrow osteoclast precursors (OCPs) in vitro. In this study, interleukin-1 receptor antagonist deficient (Il1rn−/−) and wild-type (WT) mice were compared to investigate the effects of enhanced IL-1 signaling on the composition of OCPs in long bone, calvaria, vertebra, and jaw. Bone marrow cells were isolated from these sites and the percentage of early blast (CD31hi Ly-6C−), myeloid blast (CD31+ Ly-6C+), and monocyte (CD31− Ly-6Chi) OCPs was assessed by flow cytometry. At the time-point of cell isolation, Il1rn−/− mice showed no inflammation or bone destruction yet as determined by histology and microcomputed tomography. However, Il1rn−/− mice had an approximately two-fold higher percentage of OCPs in long bone and jaw marrow compared to WT. Conversely, vertebrae and calvaria marrow contained a similar composition of OCPs in both strains. Bone marrow cells were cultured with macrophage colony stimulating factor (M-CSF) and receptor of NfκB ligand (RANKL) on bone slices to assess osteoclastogenesis and on calcium phosphate-coated plates to analyze mineral dissolution. Deletion of Il1rn increased osteoclastogenesis from long bone, calvaria, and jaw marrows, and all Il1rn−/− cultures showed increased mineral dissolution compared to WT. However, osteoclast markers increased exclusively in Il1rn−/− osteoclasts from long bone and jaw. Collectively, these findings indicate that a lack of IL-1RA increases the numbers of OCPs in vivo, particularly in long bone and jaw, where rheumatoid arthritis and periodontitis develop. Thus, increased bone loss at these sites may be triggered by a larger pool of OCPs due to the disruption of IL-1 inhibitors.

Download Full-text

Effect of Zinc Supplementation on Renal Anemia in 5/6-Nephrectomized Rats and a Comparison with Treatment with Recombinant Human Erythropoietin

International Journal of Molecular Sciences ◽

10.3390/ijms20204985 ◽

2019 ◽

Vol 20 (20) ◽

pp. 4985 ◽

Cited By ~ 2

Author(s):

Hui-Lin Feng ◽

Yen-Hua Chen ◽

Sen-Shyong Jeng

Keyword(s):

Bone Marrow ◽

Recombinant Human Erythropoietin ◽

Zinc Supplementation ◽

Bone Marrow Cells ◽

Human Erythropoietin ◽

Post Surgery ◽

Rat Bone ◽

Marrow Cells

Anemia is a severe complication in patients with chronic kidney disease (CKD). Treatment with exogenous erythropoietin (EPO) can correct anemia in many with CKD. We produced 5/6-nephrectomized rats that became uremic and anemic at 25 days post surgery. Injection of the anemic 5/6-nephrectomized rats with 2.8 mg zinc/kg body weight raised their red blood cell (RBC) levels from approximately 85% of the control to 95% in one day and continued for 4 days. We compared the effect of ZnSO4 and recombinant human erythropoietin (rHuEPO) injections on relieving anemia in 5/6-nephrectomized rats. After three consecutive injections, both the ZnSO4 and rHuEPO groups had significantly higher RBC levels (98 ± 6% and 102 ± 6% of the control) than the saline group (90 ± 3% of the control). In vivo, zinc relieved anemia in 5/6-nephrectomized rats similar to rHuEPO. In vitro, we cultured rat bone marrow cells supplemented with ZnCl2, rHuEPO, or saline. In a 4-day suspension culture, we found that zinc induced erythropoiesis similar to rHuEPO. When rat bone marrow cells were supplement-cultured with zinc, we found that zinc stimulated the production of EPO in the culture medium and that the level of EPO produced was dependent on the concentration of zinc supplemented. The production of EPO via zinc supplementation was involved in the process of erythropoiesis.

Download Full-text

Increases in circulating megakaryocyte growth-promoting activity in the plasma of rats following whole body irradiation

Blood ◽

10.1182/blood.v63.5.1060.1060 ◽

1984 ◽

Vol 63 (5) ◽

pp. 1060-1066 ◽

Cited By ~ 15

Author(s):

M Miura ◽

CW Jackson ◽

SA Lyles

Keyword(s):

Bone Marrow ◽

Plasma Concentration ◽

Bone Marrow Cells ◽

Single Cells ◽

Rat Plasma ◽

Whole Body ◽

Growth Promoting ◽

Marrow Cells

Abstract To gain insight into the regulation of megakaryocyte precursors in vivo, we assayed (in vitro) megakaryocyte growth-promoting activity (Meg-GPA) in plasma of rats in which both marrow hypoplasia and thrombocytopenia had been induced by irradiation. Rats received whole body irradiation of 834 rad from a 137Cs source. Plasma was collected at intervals of hours to days, up through day 21 postirradiation, and was tested, at a concentration of 30%, for Meg-GPA on bone marrow cells cultured in 1.1% methylcellulose with 5 X 10(-5) M 2-mercaptoethanol. With normal rat plasma, no megakaryocyte colonies (defined as greater than or equal to 4 megakaryocytes) were seen and only a few single megakaryocytes and clusters (defined as 2 or 3 megakaryocytes) were formed. Two peaks of plasma Meg-GPA were observed after irradiation. The first appeared at 12 hr, before any decrease in marrow megakaryocyte concentration or platelet count. The second occurred on days 10–14 after irradiation, after the nadir in megakaryocyte concentration and while platelet counts were at their lowest levels. A dose-response study of plasma concentration and megakaryocyte growth, using plasma collected 11 days postirradiation, demonstrated that patterns of megakaryocyte growth were related to plasma concentration; formation of single megakaryocytes was optimal over a range of 20%-30% plasma concentration, while cluster and colony formation were optimal at a plasma concentration of 30%. All forms of megakaryocyte growth were decreased with 40% plasma. There was a linear relationship between the number of bone marrow cells plated and growth of single cells, clusters, and colonies using a concentration of 30% plasma collected 11 days after irradiation. We conclude that irradiation causes time- related increases in circulating megakaryocyte growth-promoting activity. We suggest that the irradiated rat is a good model for studying the relationships between Meg-GPA and megakaryocyte and platelet concentration in vivo.

Download Full-text