scholarly journals Endocytosis and breakdown of mitochondrial malate dehydrogenase in the rat in vivo. Effects of suramin and leupeptin

1982 ◽  
Vol 208 (1) ◽  
pp. 61-67 ◽  
Author(s):  
M K Bijsterbosch ◽  
A M Duursma ◽  
J M W Bouma ◽  
M Gruber
Keyword(s):  
Bone Marrow ◽  
Malate Dehydrogenase ◽  
Kupffer Cells ◽  
Cell Types ◽  
Cell Protein ◽  
Carbon Particles ◽  
Parenchymal Cells ◽  
In Vivo Effects ◽  
Mitochondrial Malate Dehydrogenase

1. The plasma clearance of intravenously injected 125I-labelled mitochondrial malate dehydrogenase (half-life 7 min) was not influenced by previous injection of suramin and/or leupeptin (inhibitors of intralysosomal proteolysis). 2. Pretreatment with both inhibitors considerably delayed degradation of endocytosed enzyme in liver, spleen, bone marrow and kidneys. 3. The tissue distribution of radioactivity was determined at 30 min after injection, when only 3% of the dose was left in plasma. All injected radioactivity was still present in the carcass. The major part of the injected dose was found in liver (49%), spleen (5%), kidneys (13%) and bone, including marrow (11%). 4. Liver cells were isolated 15 min after injection of labelled enzyme. We found that Kupffer cells and parenchymal cells had endocytosed the enzyme at rates corresponding to 9530 and 156 ml of plasma/day per g of cell protein respectively. Endothelial cells do not significantly contribute to uptake of the enzyme. 5. Uptake by Kupffer cells was saturable, whereas uptake by parenchymal cells was not. This suggests that these cell types endocytose the enzyme via different receptors. 6. Previous injection of carbon particles greatly decreased uptake of the enzyme by liver, spleen and bone marrow.

scholarly journals Endocytosis of lactate dehydrogenase isoenzyme M4 in rats in vivo. Experiments with enzyme labelled with O-(4-diazo-3,5-di[125I]iodobenzoyl)sucrose

1982 ◽  
Vol 202 (3) ◽  
pp. 655-660 ◽  
Author(s):  
A S H De Jong ◽  
A M Duursma ◽  
J M W Bouma ◽  
M Gruber ◽  
A Brouwer ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Endothelial Cells ◽  
Lactate Dehydrogenase ◽  
Kupffer Cells ◽  
Cell Protein ◽  
Dehydrogenase Isoenzyme ◽  
Carbon Particles ◽  
In Vivo Experiments ◽  
Parenchymal Cells

1. Pig lactate dehydrogenase isoenzyme M4 was labelled with O-(4-diazo-3,5-di[125I]iodobenzoyl)sucrose and injected intravenously into rats. Previous work has shown that this label does not influence the clearance of the enzyme (half-life about 26 min) and that it is retained within the lysosomes for several hours after endocytosis and breakdown of the protein [De Jong, Bouma & Gruber (1981) Biochem. J. 198, 45-51]. 2. The distribution of the radioactivity over a large number of tissues was determined 2 h after injection. A high percentage of the injected dose was found in liver (41%), spleen (10%) and bone including marrow (21%). 3. Autoradiography indicated uptake of the enzyme mainly by Kupffer cells of the liver, by spleen macrophages and by bone marrow macrophages. 4. Liver cells were isolated 1 h after injection of the enzyme. Kupffer cells, endothelial cells and parenchymal cells were found to endocytose the enzyme at rates corresponding to 4230, 35 and 25 ml of plasma/day per g of cell protein, respectively. 5. Previous injection of carbon particles greatly reduced the uptake of the enzyme by liver and spleen, but the uptake by bone marrow was not significantly changed.

scholarly journals The interaction in vivo of transferrin and asialotransferrin with liver cells

1987 ◽  
Vol 243 (3) ◽  
pp. 715-722 ◽  
Author(s):  
T J C van Berkel ◽  
C J Dekker ◽  
J K Kruijt ◽  
H G van Eijk
Keyword(s):  
Kupffer Cells ◽  
Cell Types ◽  
Hepatic Uptake ◽  
Cell Protein ◽  
Recognition Sites ◽  
Iron Delivery ◽  
Parenchymal Cells ◽  
Mannose Receptors

Rat transferrin or asialotransferrin doubly radiolabelled with 59Fe and 125I was injected into rats. A determination of extrahepatic and hepatic uptake indicated that asialotransferrin delivers a higher fraction of the injected 59Fe to the liver than does transferrin. In order to determine in vivo the intrahepatic recognition sites for transferrin and asialotransferrin, the liver was subfractionated into parenchymal, endothelial and Kupffer cells by a low-temperature cell isolation procedure. High-affinity recognition of transferrin (competed for by an excess of unlabelled transferrin) is exerted by parenchymal cells as well as endothelial and Kupffer cells with a 10-fold higher association (expressed per mg of cell protein) to the latter cell types. In all three cell types iron delivery occurs, as concluded from the increase in cellular 59Fe/125I ratio at prolonged circulation times of transferrin. It can be calculated that parenchymal cells are responsible for 50-60% of the interaction of transferrin with the liver, 20-30% is associated with endothelial cells and about 20% with Kupffer cells. For asialotransferrin a higher fraction of the injected dose becomes associated with parenchymal cells as well as with endothelial and Kupffer cells. Competition experiments in vivo with various sugars indicated that the increased interaction of asialotransferrin with parenchymal cells is specifically inhibited by N-acetylgalactosamine whereas mannan specifically inhibits the increased interaction of asialotransferrin with endothelial and Kupffer cells. Recognition of asialotransferrin by galactose receptors from parenchymal cells or mannose receptors from endothelial and Kupffer cells is coupled to active 59Fe delivery to the cells. It is concluded that, as well as parenchymal cells, liver endothelial and Kupffer cells are also quantitatively important intrahepatic sites for transferrin and asialotransferrin metabolism, an interaction exerted by multiple recognition sites on the various cell types.

scholarly journals Uptake and degradation of human low-density lipoprotein by human liver parenchymal and Kupffer cells in culture

1991 ◽  
Vol 276 (1) ◽  
pp. 135-140 ◽  
Author(s):  
J A A M Kamps ◽  
J K Kruijt ◽  
J Kuiper ◽  
T J C Van Berkel
Keyword(s):  
Kupffer Cells ◽  
Human Liver ◽  
Low Density Lipoprotein ◽  
Ldl Receptor ◽  
Density Lipoprotein ◽  
Cell Types ◽  
Liver Cells ◽  
Cell Protein ◽  
Parenchymal Liver ◽  
Parenchymal Cells

The association with and degradation by cultured human parenchymal liver cells and human Kupffer cells of human low-density lipoprotein (LDL) was investigated in order to define, for the human situation, the relative abilities of the various liver cell types to interact with LDL. With both human parenchymal liver cells and Kupffer cells the association of LDL with the cells followed saturation kinetics which were coupled to LDL degradation. The association of LDL (per mg of cell protein) to both cell types was comparable, but the association with human Kupffer cells was much more efficiently coupled to degradation than was the case in parenchymal cells. The capacity of human Kupffer cells to degrade LDL was consequently 18-fold higher (per mg of cell protein) than that of the human parenchymal liver cells. Competition studies showed that unlabelled LDL competed efficiently with the cell association and degradation of 125I-labelled LDL with both parenchymal and Kupffer cells, while unlabelled acetyl-LDL was ineffective. The degradation of LDL by parenchymal and Kupffer cells was blocked by chloroquine and NH4Cl, indicating that it occurs in the lysosomes. Binding and degradation of LDL by human liver parenchymal cells and human Kupffer cells appeared to be completely calcium-dependent. It is concluded that the association and degradation of LDL by human Kupffer and parenchymal liver cells proceeds through the specific LDL receptor, whereas the association of LDL to Kupffer cells is more efficiently coupled to degradation. The presence of the highly active LDL receptor on human Kupffer cells might contribute significantly to LDL catabolism by human liver, especially under conditions whereby the LDL receptor on parenchymal cells is down-regulated.

scholarly journals Plasma clearance and endocytosis of cytosolic malate dehydrogenase in the rat

1983 ◽  
Vol 210 (2) ◽  
pp. 419-428 ◽  
Author(s):  
M K Bijsterbosch ◽  
A M Duursma ◽  
J M W Bouma ◽  
M Gruber
Keyword(s):  
Bone Marrow ◽  
Malate Dehydrogenase ◽  
Half Life ◽  
Cell Protein ◽  
Net Charge ◽  
Lysosomal Fraction ◽  
First Order Kinetics ◽  
Parenchymal Cells ◽  
Cytosolic Malate Dehydrogenase ◽  
Positively Charged

1. Pig heart cytosolic malate dehydrogenase was radiolabelled with O-(4-diazo-3,5-di-[125I]iodobenzoyl)sucrose and intravenously injected into rats. Enzyme activity and radioactivity were cleared from plasma identically, with first-order kinetics, with a half-life of about 30 min. 2. The tissue distribution of radioactivity was determined at 2 h after injection. All injected radioactivity was recovered from the tissues. A high percentage of the injected dose was found in liver (37%), spleen (6%) and bone including marrow (19%). 3. Radioactivity in liver and spleen increased up to 2 h after injection and subsequently declined, with a half-life of about 20 h. 4. After differential fractionation of liver, radioactivity was largely found in the mitochondrial and lysosomal fraction. 5. Liver cells were isolated 1 h after injection of labelled enzyme. We found that Kupffer cells, endothelial cells and parenchymal cells had endocytosed the enzyme at rates corresponding to 2725, 94 and 63 ml of plasma/day per g of cell protein respectively. 6. Radioautography indicated that in spleen and bone marrow the enzyme is mainly taken up by macrophages. 7. Internalization of the enzyme by liver, spleen and bone marrow was saturable. This indicates that the enzyme is taken up in these tissues by adsorptive endocytosis. 8. The present results closely resemble those obtained previously for the mitochondrial isoenzyme of malate dehydrogenase and for lactate dehydrogenase M4. Since those enzymes are positively charged at physiological pH, whereas cytosolic malate dehydrogenase is negative, net charge cannot be the major factor determining the rate of uptake of circulating enzymes by reticuloendothelial macrophages, as has been suggested in the literature [Wachsmuth & Klingmüller (1978) J. Reticuloendothel. Soc. 24, 227-241].

scholarly journals The interplay of osteogenesis and hematopoiesis

2004 ◽  
Vol 167 (6) ◽  
pp. 1113-1122 ◽  
Author(s):  
Sergei A. Kuznetsov ◽  
Mara Riminucci ◽  
Navid Ziran ◽  
Takeo W. Tsutsui ◽  
Alessandro Corsi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Parathyroid Hormone ◽  
Ex Vivo ◽  
Cell Types ◽  
Heterotopic Bone ◽  
Stromal Compartment ◽  
Marrow Space ◽  
Stromal Tissue

The ontogeny of bone marrow and its stromal compartment, which is generated from skeletal stem/progenitor cells, was investigated in vivo and ex vivo in mice expressing constitutively active parathyroid hormone/parathyroid hormone–related peptide receptor (PTH/PTHrP; caPPR) under the control of the 2.3-kb bone-specific mouse Col1A1 promoter/enhancer. The transgene promoted increased bone formation within prospective marrow space, but delayed the transition from bone to bone marrow during growth, the formation of marrow cavities, and the appearance of stromal cell types such as marrow adipocytes and cells supporting hematopoiesis. This phenotype resolved spontaneously over time, leading to the establishment of marrow containing a greatly reduced number of clonogenic stromal cells. Proliferative osteoprogenitors, but not multipotent skeletal stem cells (mesenchymal stem cells), capable of generating a complete heterotopic bone organ upon in vivo transplantation were assayable in the bone marrow of caPPR mice. Thus, PTH/PTHrP signaling is a major regulator of the ontogeny of the bone marrow and its stromal tissue, and of the skeletal stem cell compartment.

In Vivo Effects of Human Bone Marrow Mesenchymal Stromal Cells on the Development of Experimental B16 Melanoma in Mice

2020 ◽  
Vol 168 (4) ◽  
pp. 561-565 ◽  
Author(s):  
V. N. Petrov ◽  
E. V. Isaeva ◽  
S. E. Ulyanenko ◽  
E. E. Beketov ◽  
E. M. Yatsenko ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
B16 Melanoma ◽  
Human Bone ◽  
Human Bone Marrow ◽  
In Vivo Effects

scholarly journals Recombinant human granulocyte colony-stimulating factor. Effects on hematopoiesis in normal and cyclophosphamide-treated primates.

1987 ◽  
Vol 165 (4) ◽  
pp. 941-948 ◽  
Author(s):  
K Welte ◽  
M A Bonilla ◽  
A P Gillio ◽  
T C Boone ◽  
G K Potter ◽  
...  
Keyword(s):  
Bone Marrow ◽  
White Blood Cells ◽  
Chemotherapeutic Agents ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Radiation Induced ◽  
Peripheral White Blood Cells ◽  
In Vivo Effects ◽  
Stimulating Factor

We examined the in vivo effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) in primates (cynomolgus monkeys) treated with subcutaneous doses of rhG-CSF for 14-28 d. A dose-dependent increase in the peripheral white blood cells (WBC) was seen, reaching a plateau after 1 wk of rhG-CSF treatment. The elevation of WBC was due to an increase in the absolute neutrophil count. These results demonstrate that rhG-CSF is a potent granulopoietic growth and differentiation factor in vivo. In cyclophosphamide (CY)-induced myelosuppression, rhG-CSF was able to shorten the time period of WBC recovery in two treated monkeys to 1 wk, as compared to more than 4 wk for the control monkey. Its ability to significantly shorten the period of chemotherapy-induced bone marrow hypoplasia may allow clinicians to increase the frequency or dosage of chemotherapeutic agents. In addition, the increase in absolute numbers of functionally active neutrophils may have a profound effect in the rate and severity of neutropenia-related sepsis. Furthermore, the activities reported here indicate a potential role for rhG-CSF in the treatment of patients with myelodysplastic syndrome, congenital agranulocytosis, radiation-induced myelosuppression, and bone marrow transplantation.

280 MULTILINEAGE POTENTIAL OF PORCINE BONE MARROW AND ADIPOSE-DERIVED MESENCHYMAL STEM CELLS IN 3-D ALGINATE HYDROGELS

2009 ◽  
Vol 21 (1) ◽  
pp. 237 ◽  
Author(s):  
D. Kim ◽  
A. J. Maki ◽  
H.-J. Kong ◽  
E. Monaco ◽  
M. Bionaz ◽  
...  
Keyword(s):  
Gene Expression ◽  
Tissue Engineering ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Adipogenic Differentiation ◽  
Cell Types ◽  
Scaffold Material ◽  
Over Time

Adipose tissue presents an appealing alternative to bone marrow as a source of mesenchymal stem cells (MSC). However, in order to enhance cell proliferation and differentiation, 3-dimensional (3-D) culture may be required. A 3-D culture has benefits due to its more in vivo-like environment. Further, to form a functional tissue, a scaffold material is required to ensure proper shape and allow for efficient delivery of nutrients and growth factors. Alginate, a resorbable hydrogel, is a potential injectable scaffold for fat and bone tissue engineering due to its high biocompatibility, gelation with calcium and slow dissolution in a physiologic environment. In the present study, we examined the viability, gene expression and morphology of MSC, isolated from porcine adipose (ADSC) and bone marrow (BMSC), during osteogenic and adipogenic differentiation in a 3D alginate hydrogel environment for 0, 7 and 14 days (d). ADSC and BMSC were infused into alginate hydrogels, which polymerized upon the addition of Ca+2 ions. Both stem cell types were differentiated into osteoblasts using 0.1 μm dexamethasone, 10 mm beta glycerophosphate and 50 μm ascorbic acid, whereas adipocytes were differentiated using 10 μm insulin, 1 μm dexamethasone, and 0.5 mm IBMX. Osteogenic differentiation was confirmed using alkaline phosphatase, Von Kossa, and alizarin red S staining and adipogenic differentiation was confirmed using Oil Red O. Cell viability and proliferation was quantified using the MTT assay. Gene expression was measured using qPCR. The morphology of ADSC and BMSC differentiated toward osteogenic lineages changed with both cell types forming osteogenic nodules over time. The nodules formed by ADSC were larger in diameter than those formed by BMSC. Unlike the osteogenic cells that formed nodules, the ADSC and BMSC differentiated into adipogenic cells showed no significant changes in cell size or aggregation. Gene expression results indicated increased PPARG expression in BMSC with time whereas ADSC showed a peak of expression on day 7 and then decreased. ADSC showed increased (14-fold) PPRG expression when compared with BMSC. ADSC had 160-fold less expression of ALP than BMSC. BMSC showed a 16-fold higher expression level of BGLAP than ADSC. ADSC showed a 15.8% higher expression than BMSC for COL1a1. Both ADSC and BMSC showed similar trends SPARC expression, but BMSC had a 12-fold higher expression of SPP1 than ADSC. In summary, both types of mesenchymal stem cells successfully differentiated into both lineages and maintained viability in the hydrogel over time. In conclusion, alginate is a viable scaffold material for the differentiation of mesenchymal stem cells for tissue engineering applications. These results allow for future studies using the pig as an in vivo fat and bone tissue engineering model. This research was supported by the Illinois Regenerative Medicine Institute.

STAT5 promotes multilineage hematolymphoid development in vivo through effects on early hematopoietic progenitor cells

Blood ◽  
2002 ◽  
Vol 99 (1) ◽  
pp. 95-101 ◽  
Author(s):  
Jonathan W. Snow ◽  
Ninan Abraham ◽  
Melissa C. Ma ◽  
Nancy W. Abbey ◽  
Brian Herndier ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Blood Cell ◽  
Progenitor Cells ◽  
Cell Types ◽  
Hematopoietic Stem ◽  
Signal Transducers ◽  
Adult Mice ◽  
Colony Forming Units ◽  
Proliferation Activity

The transcription factor signal transducers and activators of transcription 5 (STAT5) is activated by numerous cytokines that orchestrate blood cell development. Multilineage peripheral blood cytopenias were observed in adult mice lacking both isoforms of STAT5 (STAT5A and STAT5B) as well as accelerated rates of apoptosis in the bone marrow. Although the hematopoietic stem cell (HSC) population was preserved in a number of these mice, the post-HSC progenitor populations were diminished and a marked reduction in functional progenitors (spleen colony-forming units) was detected. Competitive bone marrow transplantation studies in vivo revealed a profound impairment of repopulation potential of STAT5-null HSCs, leading to complete lack of contribution to the myeloid, erythroid, and lymphoid lineages. These abnormalities were associated with heightened proliferation activity in the HSC fraction, suggesting the action of homeostatic mechanisms to maintain sufficient levels of diverse blood cell types for viability. Thus, STAT5 normally sustains the robust hematopoietic reserve that contributes to host viability through crucial survival effects on early progenitor cells.

scholarly journals Multi-cellular 3D human primary liver cell culture elevates metabolic activity under fluidic flow

Lab on a Chip ◽  
2015 ◽  
Vol 15 (10) ◽  
pp. 2269-2277 ◽  
Author(s):  
Mandy B. Esch ◽  
Jean-Matthieu Prot ◽  
Ying I. Wang ◽  
Paula Miller ◽  
Jose Ricardo Llamas-Vidales ◽  
...  
Keyword(s):  
Cell Culture ◽  
Metabolic Activity ◽  
Kupffer Cells ◽  
Liver Cell ◽  
Low Cost ◽  
Stellate Cells ◽  
Cell Types ◽  
Liver Cell Culture ◽  
Parenchymal Cells

We have developed a low-cost liver cell culture device that creates fluidic flow over a 3D primary liver cell culture that consists of multiple liver cell types, including hepatocytes and non-parenchymal cells (fibroblasts, stellate cells, and Kupffer cells).

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