Establishment and Characterization of an Omasal Epithelial Cell Model Derived from Dairy Calves for the Study of Small Peptide Absorption
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{"title"=>"Establishment and characterization of an omasal epithelial cell model derived from dairy calves for the study of small peptide absorption", "type"=>"journal", "authors"=>[{"first_name"=>"Qingbiao", "last_name"=>"Xu", "scopus_author_id"=>"56111833400"}, {"first_name"=>"Yueming", "last_name"=>"Wu", "scopus_author_id"=>"55525387000"}, {"first_name"=>"Hongyun", "last_name"=>"Liu", "scopus_author_id"=>"53871580400"}, {"first_name"=>"Yingming", "last_name"=>"Xie", "scopus_author_id"=>"56111908200"}, {"first_name"=>"Xinbei", "last_name"=>"Huang", "scopus_author_id"=>"56112045900"}, {"first_name"=>"Jianxin", "last_name"=>"Liu", "scopus_author_id"=>"36077931900"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-84898479206", "doi"=>"10.1371/journal.pone.0088993", "sgr"=>"84898479206", "pmid"=>"24633419", "issn"=>"19326203", "pui"=>"372817381"}, "id"=>"372fda92-13c3-3519-bdc7-bb4f4da637c9", "abstract"=>"The objective of this study was to establish a primary culture of omasal epithelial cells (OECs) derived from dairy calves and to characterize its function in small peptide absorption. Bovine omasal tissues were obtained from newborn Chinese Holstein calves and digested with a 2.5% trypsin solution to obtain OECs. The isolated cells were later cultured in DMEM containing 10% fetal bovine serum, 5 μg/ml insulin, 10 ng/ml epidermal growth factor, 100 U/ml penicillin, 100 μg/ml streptomycin, 50 μg/ml gentamycin and 2.5 μg/ml amphotericin B. Hematoxylin and eosin staining of omasal tissue after digestion indicated that the cultured cells originated from the epithelial strata. Pure epithelial cells displayed an epithelial cell-like morphology, similar to cobblestone, with few visible fibroblasts and were cytokeratin 18-positive according to immunocytochemical analyses. The OECs were morphologically characterized with desmosomes, tight junctions and microvilli. These cells exhibited normal growth properties, as assessed using a cell growth curve, and were stably cultured for 10 passages. The OECs expressed the peptide transporter 1 (PEPT1) mRNA and absorbed intact glycylsarcosine (Gly-Sar). The uptake of Gly-Sar by OECs was pH-dependent with an optimal pH of 5.5-6.5. Furthermore, the uptake of Gly-Sar was also time-dependent, concentration-dependent and temperature-dependent. Moreover, PEPT1 was saturated with Gly-Sar at a concentration of 2.5 mM. The uptake via PEPT1 was higher compared with that via passive route at low substrate concentrations (<1.5 mM). This result suggested that PEPT1 contributed more to total small peptide absorption at low concentrations. In addition, this uptake could be competitively inhibited by methionine-glycine. Taken together, these data suggested that PEPT1 contributes to small peptide absorption in OECs. Thus, OECs may serve as a useful culture model for the study of the absorption of small peptides in bovine omasum.", "link"=>"http://www.mendeley.com/research/establishment-characterization-omasal-epithelial-cell-model-derived-dairy-calves-study-small-peptide", "reader_count"=>3, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Researcher"=>1, "Student > Ph. D. Student"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Researcher"=>1, "Student > Ph. D. Student"=>1}, "reader_count_by_subject_area"=>{"Agricultural and Biological Sciences"=>2, "Veterinary Science and Veterinary Medicine"=>1}, "reader_count_by_subdiscipline"=>{"Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>2}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1420380"], "description"=>"<p>A: Uptake of Gly-Sar at various pHs for 15 min. B: The uptake of Gly-Sar at various time points (min) under pH 6.5. N = 4. Values with different letter (a, b) indicate significant difference (<i>P</i><0.05).</p>", "links"=>[], "tags"=>["agriculture", "Animal management", "cell biology", "Cell processes", "Cell growth", "Cellular types", "Molecular cell biology", "anatomy", "biological tissue", "epithelium", "Epithelial cells", "Biochemistry", "metabolism", "Biological transport", "Veterinary science", "Veterinary anatomy", "Animal digestive anatomy", "ph", "gly-sar", "uptake"], "article_id"=>962189, "categories"=>["Biological Sciences"], "users"=>["Qingbiao Xu", "Yueming Wu", "Hongyun Liu", "Yingming Xie", "Xinbei Huang", "Jianxin Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0088993.g008", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_effects_of_pH_and_time_on_Gly_Sar_uptake_by_OECs_/962189", "title"=>"The effects of pH and time on Gly-Sar uptake by OECs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-14 03:18:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/1420375"], "description"=>"<p>A: A polarized monolayer was established with apical microvilli (MV) and a basal lamina on the plastic substratum; B: Connections between neighboring cells via desmosomes (De), tight junctions (TJ) and basolateral membrane infoldings (*) were visible. Tonofilaments (T) were also observed in the OECs. C and D: SEM observations revealed numerous microvilli-like structures on the surface of the OEC monolayer.</p>", "links"=>[], "tags"=>["agriculture", "Animal management", "cell biology", "Cell processes", "Cell growth", "Cellular types", "Molecular cell biology", "anatomy", "biological tissue", "epithelium", "Epithelial cells", "Biochemistry", "metabolism", "Biological transport", "Veterinary science", "Veterinary anatomy", "Animal digestive anatomy", "electron", "micrograph", "scanning", "oec"], "article_id"=>962184, "categories"=>["Biological Sciences"], "users"=>["Qingbiao Xu", "Yueming Wu", "Hongyun Liu", "Yingming Xie", "Xinbei Huang", "Jianxin Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0088993.g005", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Transmission_electron_micrograph_TEM_A_and_B_and_scanning_electron_micrograph_SEM_C_and_D_of_OEC_monolayers_/962184", "title"=>"Transmission electron micrograph (TEM, A and B) and scanning electron micrograph (SEM, C and D) of OEC monolayers.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-14 03:18:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/1420373"], "description"=>"<p>The log phase started after day 2 of the lag phase with a sharper inclination and cells entering the logarithmic growth phase on day 5. N = 4.</p>", "links"=>[], "tags"=>["agriculture", "Animal management", "cell biology", "Cell processes", "Cell growth", "Cellular types", "Molecular cell biology", "anatomy", "biological tissue", "epithelium", "Epithelial cells", "Biochemistry", "metabolism", "Biological transport", "Veterinary science", "Veterinary anatomy", "Animal digestive anatomy", "oecs"], "article_id"=>962182, "categories"=>["Biological Sciences"], "users"=>["Qingbiao Xu", "Yueming Wu", "Hongyun Liu", "Yingming Xie", "Xinbei Huang", "Jianxin Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0088993.g004", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Growth_curve_of_OECs_in_culture_/962182", "title"=>"Growth curve of OECs in culture.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-14 03:18:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/1420370"], "description"=>"<p>A: Epithelial cell suspensions obtained from newborn calves after enzymatic digestion (×200); B: Most of the omasal epithelial cells adhered to the wall of plastic substratum with small number of fibroblasts after 12 h in culture (×200); C: Cell clusters were observed after 2 d in culture (×100); D: A confluent monolayer formed after approximately one week in culture (×100); E: Purified cultures displayed a cobble-stone-like morphology (×100); F: A confluent monolayer of spindle-shaped, fibroblast-like cells (×100); Fibroblast-like cells are indicated by the arrows.</p>", "links"=>[], "tags"=>["agriculture", "Animal management", "cell biology", "Cell processes", "Cell growth", "Cellular types", "Molecular cell biology", "anatomy", "biological tissue", "epithelium", "Epithelial cells", "Biochemistry", "metabolism", "Biological transport", "Veterinary science", "Veterinary anatomy", "Animal digestive anatomy", "omasal", "epithelial", "cells"], "article_id"=>962179, "categories"=>["Biological Sciences"], "users"=>["Qingbiao Xu", "Yueming Wu", "Hongyun Liu", "Yingming Xie", "Xinbei Huang", "Jianxin Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0088993.g002", "stats"=>{"downloads"=>0, "page_views"=>20, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Morphology_of_isolated_omasal_epithelial_cells_at_different_growth_periods_/962179", "title"=>"Morphology of isolated omasal epithelial cells at different growth periods.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-14 03:18:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/1420383"], "description"=>"<p>A: OEC monolayers were incubated with various concentrations (0–5 mM) of Gly-Sar at 37°C (⧫) or 4°C (▴). Active (▪) uptake via PEPT1 was estimated according to the difference between 37°C and 4°C. B: The uptake of 0.5 mM Gly-Sar was measured in the absence or presence of increasing concentrations of methionine-glycine (0–5 mM). Uptake of Gly-Sar measured in the absence of the inhibitor dipeptide was set as 100%. N = 4.</p>", "links"=>[], "tags"=>["agriculture", "Animal management", "cell biology", "Cell processes", "Cell growth", "Cellular types", "Molecular cell biology", "anatomy", "biological tissue", "epithelium", "Epithelial cells", "Biochemistry", "metabolism", "Biological transport", "Veterinary science", "Veterinary anatomy", "Animal digestive anatomy", "substrate", "inhibitor", "gly-sar", "uptake"], "article_id"=>962192, "categories"=>["Biological Sciences"], "users"=>["Qingbiao Xu", "Yueming Wu", "Hongyun Liu", "Yingming Xie", "Xinbei Huang", "Jianxin Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0088993.g009", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_substrate_concentration_temperature_and_competitive_inhibitor_on_Gly_Sar_uptake_by_OECs_/962192", "title"=>"Effects of substrate concentration, temperature and competitive inhibitor on Gly-Sar uptake by OECs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-14 03:18:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/1420378"], "description"=>"<p>Lane L: 100 bp DNA ladder; lane 1: Omasal epithelial tissue; lane 2: OECs; lane 3 and 4: Loading control of the expression of the housekeeping gene glyceraldehydes-3-phosphate dehydrogenase (GAPDH) of tissue (3) and omasal epithelial cells (4).</p>", "links"=>[], "tags"=>["agriculture", "Animal management", "cell biology", "Cell processes", "Cell growth", "Cellular types", "Molecular cell biology", "anatomy", "biological tissue", "epithelium", "Epithelial cells", "Biochemistry", "metabolism", "Biological transport", "Veterinary science", "Veterinary anatomy", "Animal digestive anatomy", "transcription", "pcr", "pept1", "omasal", "epithelial"], "article_id"=>962187, "categories"=>["Biological Sciences"], "users"=>["Qingbiao Xu", "Yueming Wu", "Hongyun Liu", "Yingming Xie", "Xinbei Huang", "Jianxin Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0088993.g007", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Reverse_transcription_PCR_of_PEPT1_gene_expression_in_omasal_epithelial_tissue_and_OECs_/962187", "title"=>"Reverse transcription PCR of PEPT1 gene expression in omasal epithelial tissue and OECs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-14 03:18:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/1420377"], "description"=>"<p>Fluorescent images of OECs stained for cytokeratin 18 (A), PEPT1 (B) and negative control using the secondary antibody only (C). Nuclei were stained with DAPI.</p>", "links"=>[], "tags"=>["agriculture", "Animal management", "cell biology", "Cell processes", "Cell growth", "Cellular types", "Molecular cell biology", "anatomy", "biological tissue", "epithelium", "Epithelial cells", "Biochemistry", "metabolism", "Biological transport", "Veterinary science", "Veterinary anatomy", "Animal digestive anatomy", "staining"], "article_id"=>962186, "categories"=>["Biological Sciences"], "users"=>["Qingbiao Xu", "Yueming Wu", "Hongyun Liu", "Yingming Xie", "Xinbei Huang", "Jianxin Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0088993.g006", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunofluorescence_staining_of_OECs_/962186", "title"=>"Immunofluorescence staining of OECs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-14 03:18:01"}
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PMC Usage Stats | Further Information

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Relative Metric

{"start_date"=>"2014-01-01T00:00:00Z", "end_date"=>"2014-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences", "average_usage"=>[291]}, {"subject_area"=>"/Biology and life sciences/Anatomy", "average_usage"=>[267]}, {"subject_area"=>"/Biology and life sciences/Cell biology", "average_usage"=>[286]}, {"subject_area"=>"/Biology and life sciences/Molecular biology", "average_usage"=>[292, 461]}, {"subject_area"=>"/Biology and life sciences/Organisms", "average_usage"=>[310]}, {"subject_area"=>"/Medicine and health sciences", "average_usage"=>[285]}, {"subject_area"=>"/Medicine and health sciences/Anatomy", "average_usage"=>[266]}]}
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