scholarly journals Microwave fixation improves antigenicity of glutaraldehyde-sensitive antigens while preserving ultrastructural detail.

1995 ◽  
Vol 43 (3) ◽  
pp. 307-311 ◽  
Author(s):  
M C Jamur ◽  
C D Faraco ◽  
L O Lunardi ◽  
R P Siraganian ◽  
C Oliver
Keyword(s):  
Bone Marrow ◽  
Mast Cells ◽  
Plasma Membranes ◽  
Cell Types ◽  
Cell Plasma ◽  
Morphological Detail ◽  
Strong Staining ◽  
Microwave Fixation ◽  
Different Cell Types ◽  
Rat Mast Cells

Microwave fixation for electron microscopy has been used primarily for post-embedding immunocytochemistry. The present study examined the ability of microwave fixation to preserve the antigenicity of glutaraldehyde-sensitive antigens for pre-embedding immunocytochemistry. Five monoclonal antibodies (MAbs) directed against cell surface components of rat mast cells were tested. The MAbs failed to show any labeling of conventionally fixed rat bone marrow-derived mast cells even at glutaraldehyde concentrations as low as 0.1%. Strong staining of mast cell plasma membranes was seen when bone marrow was initially fixed with 2% formaldehyde and then refixed in 2% glutaraldehyde/2% formaldehyde after immunostaining. However, the ultrastructural preservation of the cells was poor. Antigenicity and morphological detail were both preserved when bone marrow was fixed in 0.05% glutaraldehyde/2% formaldehyde for 4 sec in a 550-W microwave oven. With this method, mast cells in various stages of maturation as well as cells that did not contain granules were immunoreactive. This method should prove useful with antigens from many different cell types that are sensitive to glutaraldehyde fixation.

scholarly journals The cultivation of bone marrow cells and cell lines on polymeric films

2011 ◽  
Vol 57 (5) ◽  
pp. 535-543
Author(s):  
M.S. Dolgikh ◽  
D.N. Livak ◽  
M.E. Krasheninnikov ◽  
N.A. Onishchenko
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Covalent Binding ◽  
Cell Types ◽  
Polymeric Films ◽  
Artificial Organ ◽  
Hep G2 ◽  
Different Cell Types ◽  
Cultural Media ◽  
Marrow Cells

The cultivation of multipotent mesenchymal stromal bone marrow cells and cells of A-431, MDCK, Vero, 3T3 and Hep-G2 was performed on polymeric films (PVA) with different hydrophobic fatty acid residues. The cells of different types grew on these films with different intensity, but in the most cases comparable with the cultivation control on usual plastic. The examined films were nontoxic to cells and sufficiently adhesive. They did not changed pH of cultural media, were optically transparent under microscope and comfortable in the experimental work. These films can be used as a model for the artificial organ construction. The covalent binding of different fatty acids to PVA shows possibility of the adaptable changes of films properties (hydrophobity and adhesiveness), and therefore possibility of the creation of optimal conditions for different cell types attachement and growth.

scholarly journals Continuous Free-Flow Fractionation of Cellular Constituents in Rat Bone Marrow

Blood ◽  
1965 ◽  
Vol 25 (1) ◽  
pp. 63-72 ◽  
Author(s):  
HOWARD C. MEL ◽  
LINDA T. MITCHELL ◽  
BO THORELL
Keyword(s):  
Bone Marrow ◽  
Cell Types ◽  
Single Cell Suspension ◽  
Physical Classification ◽  
Normal Rat ◽  
Good Repeatability ◽  
Different Cell Types ◽  
Stable Flow ◽  
Rat Bone

Abstract A single-cell suspension of normal rat bone marrow is prepared mechanically. This suspension is continuously fractionated in free solution, under sedimentation rate conditions, using 1 g. only. With a sample flow of 2.2 x 106 cells/minute and a 32-minute steady-state residence time in the stable-flow free boundary (STAFLO) flow-cell, the cells exit almost entirely into 7 of the 12 collection bottles. Maximum numbers of different cell types are observed, with good repeatability, in approximately descending order from top to bottom as follows: erythrocytes, "erythroblasts," "immatures," "myelocytes," and mature granulocytes. Major changes are effected relative to the starting marrow composition, and large relative enrichments are achieved for certain cell types. In addition to the rapid, mild, preparative aspect of this study, nominal sedimentation rates can be assigned for the different collection fractions, in the range of 3 x 105 to 4 x 106 svedbergs, thus making a start on this kind of simple physical classification of the cellular elements in this complex tissue.

scholarly journals The Lectin ArtinM Induces Recruitment of Rat Mast Cells from the Bone Marrow to the Peritoneal Cavity

PLoS ONE ◽  
2010 ◽  
Vol 5 (3) ◽  
pp. e9776 ◽  
Author(s):  
Patricia Andressa de Almeida Buranello ◽  
Maria Raquel Isnard Moulin ◽  
Devandir Antonio Souza ◽  
Maria Célia Jamur ◽  
Maria Cristina Roque-Barreira ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Mast Cells ◽  
Peritoneal Cavity ◽  
Rat Mast Cells

scholarly journals Immunomagnetic Isolation of Rat Bone Marrow-derived and Peritoneal Mast Cells

1997 ◽  
Vol 45 (12) ◽  
pp. 1715-1722 ◽  
Author(s):  
Maria Celia Jamur ◽  
Ana Cristina G. Grodzki ◽  
Andrea N. Moreno ◽  
William D. Swaim ◽  
Reuben P. Siraganian ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Mast Cell ◽  
Mast Cells ◽  
Cell Types ◽  
Ultrastructural Level ◽  
Immunomagnetic Separation ◽  
Specific Cell ◽  
Morphological Examination ◽  
Immunomagnetic Isolation ◽  
Rat Bone

Mast cells are difficult to purify from heterogeneous cell populations and to preserve, especially for pre-embedding immunostaining at the ultrastructural level. We have developed a technique that permits the isolation of a pure population of mast cells suitable for immunocytochemical studies. A rat mast cell-specific monoclonal antibody (MAb AA4) conjugated to tosylactivated Dynabeads 450 was used to immunomagnetically separate mast cells from rat bone marrow and peritoneal cell suspensions. Approximately 85% of the mast cells were recovered in the positive population that comprised virtually pure mast cells. After microwave fixation, morphological examination showed that the cells were intact and retained their ultrastructural detail. Mast cells in all stages of maturation were immunolabeled with a panel of antibodies after immunomagnetic separation. The combination of immunomagnetic separation followed by immunostaining should prove useful for the study of mast cell maturation and for the characterization of other specific cell types that are present in tissues in only limited numbers.

scholarly journals Fusion as the result of sperm–somatic cell interaction

Reproduction ◽  
2009 ◽  
Vol 138 (4) ◽  
pp. 679-687 ◽  
Author(s):  
M Mattioli ◽  
A Gloria ◽  
A Mauro ◽  
L Gioia ◽  
B Barboni
Keyword(s):  
Somatic Cell ◽  
Somatic Cells ◽  
Cell Types ◽  
Cell Interaction ◽  
Fluorescent Dye ◽  
Cell Plasma ◽  
Molecular Machinery ◽  
Different Cell Types ◽  
Boar Spermatozoa

The research has been designed to investigate whether acrosome-reacted spermatozoa can fuse with somatic cells and to check whether this event may involve the molecular machinery implicated in the sperm–egg fusion. Boar spermatozoa were capacitatedin vitroand then treated with A23187 to induce acrosome reaction and activate their fusogenic potential. Reacted spermatozoa, loaded with the membrane-permeant fluorescent dye calcein AM, were incubated with plated granulosa cells or cells derived from stable cell lines: CRFK, VERO, and ESK4. The fusion between spermatozoa and somatic cells was revealed by the diffusion of the fluorescent dye from the sperm to the cell as membrane fusion and cytoplasmic continuity between the two cells were established. The involvement of integrin α6 and tetraspanin CD9 in the process of fusion was assessed by carrying out the experiment in the presence of antibodies against these molecules. Moreover, the incidence of fusion displayed by the different cell types used was analyzed in relation to their content in the above molecules assessed by western blot and immunostaining. The role of CD9 was additionally investigated by using CD9-negative cells. The data presented demonstrate that boar spermatozoa can fuse with different somatic cell types derived from different species and the process requires the combined presence of both integrin and tetraspanin molecules on the cell plasma membrane.

Sur le comportement des mastocytes chez le rat en conditions d'hypo- et d'hypervitaminose A

Development ◽  
1971 ◽  
Vol 26 (3) ◽  
pp. 459-467
Author(s):  
Par A. Spreca ◽  
L. Modis ◽  
G. Conti
Keyword(s):  
Bone Marrow ◽  
Mast Cells ◽  
Vitamin A ◽  
Significant Rise ◽  
Hypervitaminosis A ◽  
Hypovitaminosis A ◽  
Rat Mast Cells

The behaviour of rat mast cells under conditions of high and low vitamin A The authors have studied the mast cells in normal rats and in rats under hypo- and hypervitaminosis A. The main results of their studies are: (a) in the skin (dermis), in the case of A hypervitaminosis, there is a significant rise in the number of the mastocytes with swelling and dcgranulation; (b) in lung and bone marrow, the number of mastocytes is increased under hypovitaminosis A and there is also blood congestion and lipomatosis of bone marrow. The authors give an interpretation of their results.

Abstract 14080: A Phase II Randomized, Double-blind, Controlled Trial of Combined Mesenchymal Stromal Cells and C-kit+ Cardiac Progenitor Cells in Ischemic Heart Failure: The CCTRN CONCERT-HF Trial

Circulation ◽  
2020 ◽  
Vol 142 (Suppl_3) ◽  
Author(s):  
Roberto Bolli ◽  
Raul Mitrani ◽  
Joshua M Hare ◽  
Carl J Pepine ◽  
Emerson C Perin ◽  
...  
Keyword(s):  
Heart Failure ◽  
Bone Marrow ◽  
Cell Therapy ◽  
Stromal Cells ◽  
Cell Types ◽  
Cardiac Progenitor Cells ◽  
Ischemic Heart Failure ◽  
Double Blind ◽  
Scar Size ◽  
Different Cell Types

Introduction: Although preclinical studies of cell delivery in models of ischemic heart failure (HF) suggest a beneficial interaction between mesenchymal stromal cells (MSCs) and c-kit+ cardiac progenitor cells (CPCs) resulting in additive therapeutic effects, no clinical trial has examined a combination of different cell types in ischemic HF. Furthermore, comparative studies of different cell products in humans are rare. CONCERT-HF (NCT02501811) is an NHLBI-sponsored, randomized, double-blind, placebo-controlled, Phase II trial of the Cardiovascular Cell Therapy Research Network (CCTRN) investigating feasibility, safety, and efficacy of MSCs and CPCs, alone and in combination, in patients with chronic ischemic HF. Objectives: To address the following questions: Is combined treatment with MSCs and CPCs feasible and safe in patients with ischemic HF? Do MSCs and CPCs, given alone or in combination, alleviate LV dysfunction, reduce scar size, improve quality of life, and/or augment functional capacity? Is either cell type more effective than the other? Is the combination of MSCs and CPCs more efficacious than MSCs alone or CPCs alone? Methods: Patients were randomized (1:1:1:1) to receive i) the combination of autologous bone marrow-derived MSCs and autologous CPCs, ii) MSCs alone, iii) CPCs alone, or iv) placebo. Target doses were 150 x 10 6 MSCs and 5 x 10 6 CPCs. All patients underwent bone marrow aspiration and right heart catheterization. Endomyocardial biopsy was performed only in the MSC + CPC and CPC alone groups; a “sham biopsy” was performed in the MSC alone and placebo groups. All patients underwent study product injection using the NOGA ® XP Mapping System and were followed for 12 months. Results: A total of 125 patients (116 M, 9 F), 62.5 ± 8.9 years old, were enrolled at 7 CCTRN centers between Nov 2016 and Oct 2018. Baseline LVEF (cardiac MRI) was 28.6 ± 6.1% with a mean scar size of 31.8 ± 10.9 g and NYHA class II (80%) or class III (15.2%). Conclusions: CONCERT-HF is the first cell therapy trial to assess a combination of different cell types and to directly compare two different cell products in patients with HF. All patients will complete follow-up by the end of June and final primary (12-month) safety and outcomes data will be available in August 2020.

TURNER'S SYNDROME WITH SECONDARY AMENORRHOEA AND SEX CHROMOSOME MOSAICISM

1964 ◽  
Vol 46 (3) ◽  
pp. 341-351 ◽  
Author(s):  
D. R. London ◽  
N. H. Kemp ◽  
R. Ellis ◽  
Ursula Mittwoch
Keyword(s):  
Bone Marrow ◽  
X Chromosome ◽  
Sex Chromosome ◽  
Acrocentric Chromosome ◽  
Ring Chromosome ◽  
Cell Types ◽  
Abdominal Skin ◽  
Chromosome Mosaicism ◽  
Different Cell Types ◽  
Bone Marrow Blood

ABSTRACT A female patient is described aged 28 years, height 145 cm, with infantile genitalia, infantile uterus and atrophic ovaries and in whom menstruation had occurred over a period of five years. Chromosome studies from bone marrow, blood, skin (arm and abdominal wall) and both ovaries revealed sex chromosome mosaicism and a structurally abnormal X chromosome. Three cell lines were observed. The prevalent cell line which was present in cultures from all tissues had 45 chromosomes and an XO karyotype; cultures from all tissues except the abdominal skin contained cells with 46 chromosomes, with an X/deleted X karyotype (the latter in the form of a large acrocentric chromosome); lastly a small dot-like (ring?) chromosome was present as the 46th chromosome in some cells derived from the abdominal skin and right ovary. The frequency of the different cell types in cultures from the ovaries differed considerably from those of other tissues.

scholarly journals Association of hyaluronan with rat vascular endothelial and smooth muscle cells.

1995 ◽  
Vol 43 (7) ◽  
pp. 689-697 ◽  
Author(s):  
P S Eggli ◽  
W Graber
Keyword(s):  
Smooth Muscle ◽  
Plasma Membranes ◽  
Vascular Tissue ◽  
Cell Types ◽  
Vascular Endothelial ◽  
Cell Plasma ◽  
One Step ◽  
Lowicryl K4m ◽  
20 Nm ◽  
Hyaluronan Binding

We investigated the distribution of hyaluronan (hyaluronic acid) in rat vascular tissue fixed by an osmium tetroxide (or glutaraldehyde) microwave technique and embedded in epoxy resin (or Lowicryl K4M), using hyaluronan binding proteins coupled to 15-20-nm gold particles as ultrastructural markers in a one-step post-embedding procedure. The intra- and extracellular aspects of vascular endothelial and smooth muscle cell plasma membranes revealed distinct labeling, with a high affinity for caveolae being manifested in both cell types and in all kinds of vessels. Hyaluronan was also localized intercellularly in areas characterized by extensive endothelial cell interdigitation. Intracellularly, moderate staining of nuclear heterochromatin was observed.

An Electron Microscopic Study of Normal Blood and Bone Marrow in Pigs

1968 ◽  
Vol 5 (5) ◽  
pp. 451-470 ◽  
Author(s):  
Per H. J. Nafstad ◽  
Inger Nafstad
Keyword(s):  
Bone Marrow ◽  
Internal Structure ◽  
Animal Species ◽  
Bone Marrow Cells ◽  
Cell Types ◽  
Immature Stages ◽  
Electron Microscopic ◽  
Irregular Shapes ◽  
Microscopic Picture ◽  
Different Cell Types

The ultrastructure of blood and bone marrow cells in the normal pig was studied; the process of maturation of the different cell types was found to be essentially in accordance with that in other animal species. The eosinophilic granules had a pattern which differed from that in other mammals, being characterized by specific internal structure in the immature stages. During maturation, however, a homogenous appearance supervened. Moreover, the lymphocytic nuclei were found to have irregular shapes as compared with the light microscopic picture. The results are compared to some reported studies of other animal species.

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