Cytoarchitecture of cultured olfactory neuroepithelial spheres: The role of olfactory sustentacular cells in cavity formation

When newborn rat olfactory mucosal cells were dissaggregated and plated as single cells onto astrocyte bed layers, abundant generation and maturation of olfactory receptor neurons was observed. Both immature and mature neurons were found in large multicellular spheres after 15 days in culture,. In sections of the spheres, almost all had central cavities, and the neurons were located almost exclusively in the external 1-2 cell layers. We show here that the spheres also contain olfactory sustentacular cells, neuronal supporting cells similar to retinal Muller cells. Sustentacular cell presence was shown by immunostaining cultured cells with a monoclonal antibody, SUS, which has previously been shown to recognize only sustentacular cells and a subset of the Bowman's gland cells. Fig. 1 shows that cultured cellular spheres contained numerous SUSpositive, roughly bipolar cells. These cells appear to be located internally rather than on the external surface. Sustentacular cell presence was further confirmed by immunostaining with a monoclonal antibody, 1F4, developed in this laboratory.

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IN VITRO METHODS FOR THE STUDY OF THE ADENOHYPOPHYSIAL FUNCTIONS TO SECRETE GONADOTROPHIN

Acta Endocrinologica ◽

10.1530/acta.0.068s027 ◽

1971 ◽

Vol 68 (1_Suppl) ◽

pp. S27-S40 ◽

Cited By ~ 1

Author(s):

T. Kobayashi ◽

T. Kigawa ◽

M. Mizuno ◽

T. Watanabe

Keyword(s):

Cell Culture ◽

Organ Culture ◽

Cultured Cells ◽

Cell Sheet ◽

Short Term ◽

Hypothalamic Extract ◽

Numerous Cell ◽

Single Cell Culture ◽

Almost All

ABSTRACT There are several in vitro methods to analyse the function of the adenohypophysis or the mechanisms of its regulation. The present paper deals with single cell culture, organ culture and short term incubation techniques by which the morphology and gonadotrophin-secreting function of the adenohypophysis were studied. In trypsin-dispersed cell culture, the adenohypophysial cells showed extensive propagation to form numerous cell colonies and finally develop into a confluent monolayer cell sheet covering completely the surface of culture vessels. Almost all of the cultured cells, however, became chromophobic, at least at the end of the first week of cultivation, when gonadotrophin was detectable neither in the culture medium nor in the cells themselves. After the addition of the hypothalamic extract, gonadotrophin became detectable again, and basophilic or PAS-positive granules also reappeared within the cells, suggesting that the gonadotrophs were stimulated by the extract to produce gonadotrophin. In organ culture and short term incubation, the incorporation of [3H] leucine into the adenohypophysial cells in relation to the addition of hypothalamic extract was examined. It was obvious that the ability to incorporate [3H] leucine into the gonadotrophs in vitro was highly dependent upon the presence of the hypothalamic extract.

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Extended haplotype-phasing of long-read de novo genome assemblies using Hi-C

Nature Communications ◽

10.1038/s41467-020-20536-y ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Zev N. Kronenberg ◽

Arang Rhie ◽

Sergey Koren ◽

Gregory T. Concepcion ◽

Paul Peluso ◽

...

Keyword(s):

Zebra Finch ◽

Cultured Cells ◽

De Novo ◽

Single Cells ◽

Variant Calling ◽

Chromatin Interaction ◽

Extended Haplotype ◽

Benchmark Datasets ◽

And Performance ◽

Genome Assemblies

AbstractHaplotype-resolved genome assemblies are important for understanding how combinations of variants impact phenotypes. To date, these assemblies have been best created with complex protocols, such as cultured cells that contain a single-haplotype (haploid) genome, single cells where haplotypes are separated, or co-sequencing of parental genomes in a trio-based approach. These approaches are impractical in most situations. To address this issue, we present FALCON-Phase, a phasing tool that uses ultra-long-range Hi-C chromatin interaction data to extend phase blocks of partially-phased diploid assembles to chromosome or scaffold scale. FALCON-Phase uses the inherent phasing information in Hi-C reads, skipping variant calling, and reduces the computational complexity of phasing. Our method is validated on three benchmark datasets generated as part of the Vertebrate Genomes Project (VGP), including human, cow, and zebra finch, for which high-quality, fully haplotype-resolved assemblies are available using the trio-based approach. FALCON-Phase is accurate without having parental data and performance is better in samples with higher heterozygosity. For cow and zebra finch the accuracy is 97% compared to 80–91% for human. FALCON-Phase is applicable to any draft assembly that contains long primary contigs and phased associate contigs.

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Detection and enumeration of colonic mucosal cells in amniotic fluid using a colon epithelial-specific monoclonal antibody

Prenatal Diagnosis ◽

10.1002/pd.1970101106 ◽

1990 ◽

Vol 10 (11) ◽

pp. 725-732 ◽

Cited By ~ 3

Author(s):

David Chitayat ◽

Robert W. Marion ◽

Linda Squillante ◽

Dagmar K. Kalousek ◽

Kiron M. Das

Keyword(s):

Monoclonal Antibody ◽

Amniotic Fluid ◽

Specific Monoclonal Antibody ◽

Mucosal Cells

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Distribution of hyaluronan in the epiphysial growth plate: turnover by CD44-expressing osteoprogenitor cells

Journal of Cell Science ◽

10.1242/jcs.107.10.2669 ◽

1994 ◽

Vol 107 (10) ◽

pp. 2669-2677 ◽

Cited By ~ 1

Author(s):

P. Pavasant ◽

T.M. Shizari ◽

C.B. Underhill

Keyword(s):

Monoclonal Antibody ◽

Growth Plate ◽

Cultured Cells ◽

Human Cell Line ◽

Histochemical Staining ◽

Long Bone ◽

Thin Sections ◽

Osteoprogenitor Cells ◽

Double Label ◽

Mouse Tibia

In the present study, we have examined the distribution of both hyaluronan and its receptor, CD44, during the process of endochondral ossification in the mouse tibia. Histochemical staining revealed that a large amount of hyaluronan was present in the lacunae located in the zone of hypertrophy, but it was greatly reduced or absent from the zone of erosion. In addition, hyaluronan was present in the cytoplasm of osteoprogenitor cells located in the zone of erosion. These cells also expressed CD44 on their surfaces, as revealed by double-label immunohistochemistry. These results suggested that the osteoprogenitor cells may use CD44 to bind and internalize hyaluronan, and subsequently degrade it with lysosomal enzymes. To test this possibility, we examined the human cell line, MG-63, which closely resembles osteoprogenitor cells. These cells produced several different forms of CD44, as determined by western blotting (85, 116 and 150 kDa). In addition, the binding of isotopically labeled hyaluronan to detergent extracts of these cells was blocked by a monoclonal antibody to CD44. Similarly, the degradation of hyaluronan by these cultured cells was also inhibited by a monoclonal antibody to CD44. To determine if these cells could remove hyaluronan from the growth plate, the cells were cultured directly on top of thin sections of the epiphysial region of long bone. After 16 hours, the sections were stained for hyaluronan. The MG-63 cells removed significant amounts of hyaluronan present in the zone of hypertrophy, and this effect was blocked by an excess of soluble hyaluronan and by a monoclonal antibody to CD44.(ABSTRACT TRUNCATED AT 250 WORDS)

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Characterization of a factor that stimulates hydrolysis of GTP bound to ras gene product p21 (GTPase-activating protein) and correlation of its activity to cell density

Molecular and Cellular Biology ◽

10.1128/mcb.8.10.4169-4173.1988 ◽

1988 ◽

Vol 8 (10) ◽

pp. 4169-4173

Author(s):

M Hoshino ◽

M Kawakita ◽

S Hattori

Keyword(s):

Gene Product ◽

Cell Density ◽

Cultured Cells ◽

Specific Activity ◽

Gtpase Activating Protein ◽

Ras Gene ◽

Cell Extracts ◽

Mouse Tissues ◽

Almost All ◽

Hydrolysis Of

The postmicrosomal fraction of the extract from NIH 3T3 and BALB/c 3T3 cells stimulated the hydrolysis of GTP bound to H-ras gene product p21 by severalfold. The stimulation was observed with normal p21 but not with p21 with valine as the 12th residue. This specificity is similar to that of GTPase-activating protein (GAP) for N-ras p21 described by M. Trahey and F. McCormick (Science 238:542-545, 1987). Consistent with this specificity, analysis of p21-bound nucleotides in living cells revealed that almost all normal p21 bound GDP, whereas oncogenic mutant p21s bound both GTP and GDP. Similar activity was also found in various mouse tissues, with brain tissue showing the highest specific activity. When cell extracts were prepared from cultured cells, there was a linear relationship between GAP activity and cell density. These results suggest the factor is involved in the regulation of cell proliferation.

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Primary culture of secretagogue-responsive parietal cells from rabbit gastric mucosa

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.1989.256.1.g254 ◽

1989 ◽

Vol 256 (1) ◽

pp. G254-G263 ◽

Cited By ~ 20

Author(s):

C. S. Chew ◽

M. Ljungstrom ◽

A. Smolka ◽

M. R. Brown

Keyword(s):

Gastric Mucosa ◽

Primary Culture ◽

Cultured Cells ◽

Single Cells ◽

Parietal Cells ◽

Ultrastructural Changes ◽

Long Term Effects ◽

H2 Receptor Antagonist ◽

Morphological Transformations

A new procedure for isolation and primary culture of gastric parietal cells is described. Parietal cells from rabbit gastric mucosa are enriched to greater than 95% purity by combining a Nycodenz gradient separation with centrifugal elutriation. Cells are plated on the basement membrane matrix, Matrigel, and maintained in culture for at least 1 wk. Parietal cells cultured in this manner remain differentiated, cross-react with monoclonal H+-K+-ATPase antibodies, and respond to histamine, gastrin, and cholinergic stimulation with increased acid production as measured by accumulation of the weak base, [14C]aminopyrine. When stimulated, cultured cells undergo ultrastructural changes in which intracellular canaliculi expand and numerous microvilli are observed. These ultrastructural changes are similar to those previously found to occur in vivo and in acutely isolated parietal cells. Morphological transformations in living cells can also be observed with differential interference contrast optics in the light microscope. After histamine stimulation, intracellular canaliculi gradually expand to form large vacuolar spaces. When the H2 receptor antagonist, cimetidine, is added to histamine-stimulated cells, these vacuoles gradually disappear. The ability to maintain hormonally responsive parietal cells in primary culture should make it possible to study direct, long-term effects of a variety of agonists and antagonists on parietal cell secretory-related activity. These cultured cells should also prove to be useful for the study of calcium transients, ion fluxes, and intracellular pH as related to acid secretion in single cells, particularly since morphological transformations can be used to monitor "physiological" responses at the same time within the same cell.

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A 2.8-Angstrom-Resolution Cryo-Electron Microscopy Structure of Human Parechovirus 3 in Complex with Fab from a Neutralizing Antibody

Journal of Virology ◽

10.1128/jvi.01597-18 ◽

2018 ◽

Vol 93 (4) ◽

Cited By ~ 5

Author(s):

Aušra Domanska ◽

Justin W. Flatt ◽

Joonas J. J. Jukonen ◽

James A. Geraets ◽

Sarah J. Butcher

Keyword(s):

Electron Microscopy ◽

Monoclonal Antibody ◽

High Resolution ◽

Cultured Cells ◽

Human Monoclonal Antibody ◽

Neutralizing Antibody ◽

Molecular Diagnostic ◽

Diagnostic Tools ◽

Human Parechovirus ◽

Cryo Electron Microscopy

ABSTRACTHuman parechovirus 3 (HPeV3) infection is associated with sepsis characterized by significant immune activation and subsequent tissue damage in neonates. Strategies to limit infection have been unsuccessful due to inadequate molecular diagnostic tools for early detection and the lack of a vaccine or specific antiviral therapy. Toward the latter, we present a 2.8-Å-resolution structure of HPeV3 in complex with fragments from a neutralizing human monoclonal antibody, AT12-015, using cryo-electron microscopy (cryo-EM) and image reconstruction. Modeling revealed that the epitope extends across neighboring asymmetric units with contributions from capsid proteins VP0, VP1, and VP3. Antibody decoration was found to block binding of HPeV3 to cultured cells. Additionally, at high resolution, it was possible to model a stretch of RNA inside the virion and, from this, identify the key features that drive and stabilize protein-RNA association during assembly.IMPORTANCEHuman parechovirus 3 (HPeV3) is receiving increasing attention as a prevalent cause of sepsis-like symptoms in neonates, for which, despite the severity of disease, there are no effective treatments available. Structural and molecular insights into virus neutralization are urgently needed, especially as clinical cases are on the rise. Toward this goal, we present the first structure of HPeV3 in complex with fragments from a neutralizing monoclonal antibody. At high resolution, it was possible to precisely define the epitope that, when targeted, prevents virions from binding to cells. Such an atomic-level description is useful for understanding host-pathogen interactions and viral pathogenesis mechanisms and for finding potential cures for infection and disease.

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The Treasury of Wharton's Jelly

Stem Cell Reviews and Reports ◽

10.1007/s12015-021-10217-8 ◽

2021 ◽

Author(s):

Rebecca Guenther ◽

Stephan Dreschers ◽

Jessika Maassen ◽

Daniel Reibert ◽

Claudia Skazik-Voogt ◽

...

Keyword(s):

Stem Cells ◽

Umbilical Cord ◽

Cultured Cells ◽

Single Cells ◽

Mesenchymal Progenitor Cells ◽

Differentiation Potential ◽

Lineage Differentiation ◽

Umbilical Cord Tissue ◽

The Common ◽

Culture Supernatants

Abstract Background Postnatal umbilical cord tissue contains valuable mesenchymal progenitor cells of various differentiation stages. While mesenchymal stem cells are plastic-adherent and tend to differentiate into myofibroblastic phenotypes, some round cells detach, float above the adherent cells, and build up cell aggregates, or form spheroids spontaneously. Very small luminescent cells are always involved as single cells or within collective forms and resemble the common well-known very small embryonic-like cells (VSELs). In this study, we investigated these VSELs-like cells in terms of their pluripotency phenotype and tri-lineage differentiation potential. Methods VSELs-like cells were isolated from cell-culture supernatants by a process that combines filtering, up concentration, and centrifugation. To determine their pluripotency character, we measured the expression of Nanog, Sox-2, Oct-4, SSEA-1, CXCR4, SSEA-4 on gene and protein level. In addition, the cultured cells derived from UC tissue were examined regarding their potential to differentiate into three germ layers. Result The VSELs-like cells express all of the pluripotency-associated markers we investigated and are able to differentiate into meso- endo- and ectodermal precursor cells. Conclusions Umbilical cord tissue hosts highly potent VSELs-like stem cells. Graphical Abstract

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Vasculogenesis in the early quail blastodisc as studied with a monoclonal antibody recognizing endothelial cells

Development ◽

10.1242/dev.100.2.339 ◽

1987 ◽

Vol 100 (2) ◽

pp. 339-349 ◽

Cited By ~ 27

Author(s):

L. Pardanaud ◽

C. Altmann ◽

P. Kitos ◽

F. Dieterlen-Lievre ◽

C.A. Buck

Keyword(s):

Monoclonal Antibody ◽

Endothelial Cells ◽

Single Cells ◽

Primitive Streak ◽

Vascular Network ◽

Vascular Tree ◽

Somite Stage ◽

Area Vasculosa ◽

Haemopoietic Cells ◽

Area Pellucida

QH1, a monoclonal antibody that recognizes quail endothelial and haemopoietic cells, was applied to quail blastodiscs in toto, in order to analyse by immunofluorescence the emergence of the vascular tree. The first endothelial cells were detected in the area opaca at the headfold stage and in the area pellucida at the 1-somite stage. Single cells then interconnected progressively, especially in the anterior intestinal portal and along the somites building up the linings of the heart and dorsal aortas. This study demonstrates that endothelial cells differentiate as single entities 4 h earlier in development than hitherto detected and that the vascular network forms secondarily. The horseshoe shape of the extraembryonic area vasculosa is also a secondary acquisition. A nonvascularized area persists until later (at least the 14-somite stage) in the region of the regressing primitive streak.

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