Role of the cellular matrix in haemopoiesis. I. Synthesis of glycosaminoglycans by mouse bone marrow cell cultures

1983 ◽  
Vol 63 (1) ◽  
pp. 155-171
Author(s):  
J.T. Gallagher ◽  
E. Spooncer ◽  
T.M. Dexter
Keyword(s):  
Bone Marrow ◽  
Hyaluronic Acid ◽  
Chondroitin Sulphate ◽  
Close Association ◽  
Heparan Sulphate ◽  
Calf Serum ◽  
Gel Chromatography ◽  
Mouse Bone Marrow ◽  
Foetal Calf Serum ◽  
Mouse Bone Marrow Cell

Haemopoietically active mouse bone marrow cultures, incubated for 48 h with [3H]glucosamine and Na2(35)SO4, synthesized radiolabelled hyaluronic acid, heparan sulphate and chondroitin sulphate. Heparan sulphate was enriched in a trypsin extract of the adherent cells whereas hyaluronic acid and chondroitin sulphate were distributed mainly to the culture medium. Analysis of nitrous acid scission products of heparan sulphate by gel chromatography demonstrated the close association of N- and O-sulphate groups along the polysaccharide chain. Chondroitinase AC degradation established the copolymeric nature of chondroitin sulphate in which about 38% of the hexuronic acid residues were in the form of GlcUA. Studies on non-haemopoietic cultures, derived from W/Wv mice or from normal marrow cells maintained in foetal calf serum instead of horse serum, indicated that adherent stromal cells were the major source of glycosaminoglycans.

Structural, luminescence and geno/cytoxicity study of carbon dots derived from Opuntia ficus-indica (L.) Mill

2020 ◽  
Vol 44 (3) ◽  
pp. 942-950
Author(s):  
Eduardo Madrigal-Bujaidar ◽  
Genaro Ivan Cerón-Montes ◽  
Joan Reyes-Miranda ◽  
Erasto Vergara-Hernández ◽  
Isela Álvarez-González ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Proliferation ◽  
Bone Marrow Cell ◽  
Carbon Dots ◽  
Marrow Cell ◽  
Mouse Bone Marrow ◽  
Opuntia Ficus Indica ◽  
Mouse Bone Marrow Cell

Carbon dots derived from nopal significantly increase the number of micronuclei in mouse erythrocytes and inhibit mouse bone marrow cell proliferation.

scholarly journals Dry Mass Distribution Changes in Mouse Bone Marrow Cell Population during the First 24 Hours after X—Irradiation as Determined by Interfereometry

Blood ◽  
1965 ◽  
Vol 25 (3) ◽  
pp. 299-309 ◽  
Author(s):  
HUN LEE ◽  
VICTOR RICHARDS ◽  
MARIA MAICHLE
Keyword(s):  
Bone Marrow ◽  
Mass Distribution ◽  
Bone Marrow Cells ◽  
Marrow Cell ◽  
Dry Mass ◽  
Steady Increase ◽  
Mouse Bone Marrow ◽  
Proliferating Cells ◽  
Mouse Bone Marrow Cell ◽  
Total Body

Abstract LAF1 mice were treated with a total-body dose of 800 r. x-ray. The dry mass distribution of the femur bone marrow cells was determined at different intervals postirradiation. The nonproliferating cells showed no significant dry mass change, whereas the proliferating cells of the myelocyte series had a steady increase in mean dry mass per cell. Few cells with the maximum dry mass increase survived at the end of 48 hours postirradiation. The dry mass distribution formed characteristic patterns for each postirradiation interval studied as the proliferating cells shifted to higher dry mass values with accompanied increase in cell sizes.

scholarly journals Highly restricted expression of a stromal cell determinant in mouse bone marrow in vivo.

1992 ◽  
Vol 176 (4) ◽  
pp. 927-935 ◽  
Author(s):  
K Jacobsen ◽  
K Miyake ◽  
P W Kincade ◽  
D G Osmond
Keyword(s):  
Bone Marrow ◽  
Stromal Cells ◽  
Stromal Cell ◽  
Plasma Membranes ◽  
Cell Clone ◽  
Close Association ◽  
Lymphoid Cells ◽  
Mouse Bone Marrow ◽  
Hemopoietic Cells

B lymphocyte precursor cells in mouse bone marrow develop in close association with stromal cells which provide essential growth signals. To identify molecules that may normally play a role in this interaction we have examined the in vivo binding of a new monoclonal antibody (mAb) (KMI6) that recognizes a determinant on a bone marrow stromal cell line (BMS2) in vitro. Flow cytometric and radioautographic evaluations revealed that the antigen recognized by KMI6 is represented on the surface of an extremely small number of cells in bone marrow cell suspensions from adult mice. An apparent molecular mass of 110 kD was obtained by surface labeling of a stromal cell clone and immunoprecipitation. Purified mAb KMI6 labeled with 125I was then given intravenously to young C3H/HeJ mice. Unbound mAb was washed out by cardiac perfusion and femoral bone marrow was examined by light and electron microscope radioautography. KMI6 labeling was heavy on the plasma membrane of many stromal cells, especially those located towards the outer subosteal region. The KMI6-labeled stromal cells were usually associated with cells of lymphoid morphology which they often completely surrounded. The labeling was restricted to areas of stromal cell plasma membranes in contact with lymphoid cells. The lymphoid cells themselves, as well as macrophages and other hemopoietic cells, failed to bind mAb KMI6 significantly. Stromal cells in bone marrow depleted of hemopoietic cells by gamma-irradiation (9,5 Gy) bound mAb KMI6 at reduced intensity. The results demonstrate that the KMI6 determinant, a 110-kD protein, is expressed on bone marrow stromal cells in vivo. Its restriction to areas of interaction with lymphoid cells suggests a role in forming microenvironmental niches of B lymphopoiesis. The surface membrane of individual stromal cells may thus be functionally polarized towards interacting B cell precursors and other hemopoietic cells.

scholarly journals Fractionation and characterization of proteoglycans isolated from chondrocyte cell cultures

1981 ◽  
Vol 197 (2) ◽  
pp. 249-258 ◽  
Author(s):  
Sven Björnsson ◽  
Dick Heinegȧrd
Keyword(s):  
Cell Culture ◽  
Chondroitin Sulphate ◽  
Calf Serum ◽  
Void Volume ◽  
Cartilage Tissue ◽  
Foetal Calf Serum ◽  
Nasal Cartilage ◽  
Core Proteins ◽  
Minor Portion ◽  
A Minor

Chondrocyte cultures were established from foetal bovine tracheal cartilage and maintained in Ham's F12 medium with or without 10% (v/v) foetal calf serum. The proteoglycans were isolated and characterized. (1) The proteoglycans from cultures both with and without serum distributed in associative or dissociative CsCl gradients like proteoglycans from cartilage tissue. (2) The amino acid composition, protein contents and glucosamine/galactosamine ratios were grossly identical with those of the tissue derived proteoglycans. (3) Sedimentation coefficients (s0) for the monomers were 21.0S and 22.7S from cultures without and with serum respectively. The s0 values obtained for aggregates were 72.3S and 93.2S respectively. The limiting viscosity numbers [η] were 248ml/g and 298ml/g respectively. These data corresponded well to those obtained for the tissue-derived proteoglycans. (4) The sizes of the core proteins and chondroitin sulphate chains respectively were the same for both types of cell-culture proteoglycans and similar to those of the tissue proteoglycans. Both the keratan sulphate-rich region and the hyaluronic acid-binding region were identified. The latter, however, was not resistant to limit digestion with trypsin, in contrast with the fragment derived from the bovine nasal cartilage. (5) About 70% of the cell-culture proteoglycans chromatographed in the void volume on a Sepharose 2B column, whereas reduced and alkylated samples (monomers) chromatographed completely included in the column. The two link proteins present in A1 preparations of cartilage proteoglycans were also present in A1 preparations of cell-culture proteoglycans. (6) A minor portion (10%) of the 35S-labelled proteoglycans in the cultures was associated with the cells. Reduced and alkylated samples were larger compared with the monomers in the medium, and chromatographed partly (25%) excluded on the Sepharose 2B column. A larger proportion (50%) of the non-reduced samples chromatographed in the void volume of the column.

scholarly journals Cell Cycle Analysis and Synchronization of Pluripotent Hematopoietic Progenitor Stem Cells

Blood ◽  
1997 ◽  
Vol 90 (6) ◽  
pp. 2293-2299 ◽  
Author(s):  
G. Prem Veer Reddy ◽  
Cheryl Y. Tiarks ◽  
Lizhen Pang ◽  
Joanne Wuu ◽  
Chung-Cheng Hsieh ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Bone Marrow ◽  
Hematopoietic Stem Cells ◽  
Calf Serum ◽  
S Phase ◽  
G1 Phase ◽  
Proliferative Potential ◽  
Mouse Bone Marrow ◽  
Hematopoietic Stem

Abstract Hematopoietic stem cells purified from mouse bone marrow are quiescent with less than 2% of Lin− Hoechstlow/Rhodaminelow (Lin− Holow/Rholow) and 10% to 15% of Lin−/Sca+ cells in S phase. These cells enter proliferative cycle and progress through G1 and into S phase in the presence of cytokines and 5% heat-inactivated fetal calf serum (HI-FCS). Cytokine-stimulated Lin− Holow/Rholow cells took 36 to 40 hours to complete first division and only 12 hours to complete each of 5 subsequent divisions. These cells require 16 to 18 hours to transit through G0 /G1 period and 28 to 30 hours to enter into mid-S phase during the first cycle. Up to 56% of Lin− Rholow/Holow cells are high-proliferative potential (7 factor-responsive) colony-forming cells (HPP-CFC). At isolation, HPP-CFC are quiescent, but after 28 to 30 hours of culture, greater than 60% are in S phase. Isoleucine-deprivation of Lin−Holow/Rholow cells in S phase of first cycle reversibly blocked them from entering into second cycle. After the release from isoleucine-block, these cells exhibited a G1 period of less than 2 hours and entered into mid-S phase by 12 hours. Thus, the duration of G1 phase of the cells in second cycle is 4 to 5 times shorter than that observed in their first cycle. Similar cell cycle kinetics are observed with Lin−/Sca+ population of bone marrow cells. Stem cell factor (SCF ) alone, in the presence of HI-FCS, is as effective as a cocktail of 2 to 7 cytokines in inducing quiescent Lin−/Sca+ cells to enter into proliferative cycle. Aphidicolin treatment reversibly blocked cytokine-stimulated Lin−/Sca+ cells at G1 /S boundary, allowing their tight synchrony as they progress through first S phase and enter into second G1 . For these cells also, SCF alone is sufficient for their progression through S phase. These studies indicate a very short G1 phase for stem cells induced to proliferate and offer experimental approaches to synchronize murine hematopoietic stem cells.

scholarly journals Expression and role of mannose receptor/terminal high-mannose type oligosaccharide on osteoclast precursors during osteoclast formation

2003 ◽  
Vol 176 (2) ◽  
pp. 285-292 ◽  
Author(s):  
S Morishima ◽  
I Morita ◽  
T Tokushima ◽  
H Kawashima ◽  
M Miyasaka ◽  
...  
Keyword(s):  
Cell Culture ◽  
Bone Marrow ◽  
Osteoclast Differentiation ◽  
Mannose Receptor ◽  
Osteoclast Formation ◽  
Tartrate Resistant Acid Phosphatase ◽  
Mouse Bone Marrow ◽  
Mouse Bone Marrow Cell ◽  
Raw264.7 Cell ◽  
High Mannose

Osteoclasts are formed from hematopoietic precursors via cell-cell fusion. We have previously reported that mannose residues are expressed on the outer membranes of monocytes during osteoclast differentiation. In the present study, we have attempted to demonstrate the pattern of expression levels of terminal high-mannose type oligosaccharide and to show that the mannose receptor is expressed on osteoclast precursor cells. Osteoclasts were formed using three different systems, namely mouse bone marrow cell culture, co-culture of mouse spleen cells with stromal cells, and RAW264.7 cell cultures. During osteoclast differentiation, the expression of terminal high-mannose type oligosaccharide gradually increased and then peaked at the stage of fusion in all three systems. Expression of the mannose receptor gradually increased during osteoclast differentiation in bone marrow cells and the co-culture system. In contrast, that in RAW264.7 cells had already been detected in the absence of the soluble receptor activator of NF-kappaB ligand and did not change during osteoclast differentiation. To ascertain whether expression of high-mannose type oligosaccharide is involved in tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cell (MNC) formation, glycosidase inhibitors were used on RAW264.7 cell culture. Castanospermine, an inhibitor of glucosidase I, inhibited the TRAP-positive MNCs, and deoxymannojirimycin, an inhibitor of alpha-mannosidase I, increased the TRAP-positive MNC formation. These results indicate that the binding of terminal high-mannose and mannose receptor is important for the process of cellular fusion in osteoclast formation.

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