In vitro cellular & developmental biology : journal of the Tissue Culture Association最新文献

{"title":"Crude liver membrane fractions as substrate preserve liver-specific functions in long-term, serum-free rat hepatocyte cultures.","authors":"B Saad,&nbsp;H Schawalder,&nbsp;P Maier","doi":"10.1007/BF02634369","DOIUrl":"https://doi.org/10.1007/BF02634369","url":null,"abstract":"<p><p>Over time, rat hepatocytes cultured on collagen lose the capacity to express liver-specific functions. The influence on this degradation process of an alternative substratum--crude membrane fractions prepared from the liver of the same rat strain--was investigated. Freshly isolated rat hepatocytes were cultured in serum-free Williams E medium supplemented with aprotinin, selenium, dexamethasone, and insulin in flasks coated with a mixture of rat liver crude membrane fractions:collagen type I (100:1). The cells adhered firmly, exhibiting minimal spreading and remaining grouped in columns or in cell islands, and retained their liver-specific functions for more than 1 wk. Hepatocytes secreted substantially higher amounts of albumin than cells cultured on collagen-coated dishes, and on Days 1 and 9 in culture the total P-450 content was 72 and 40%, respectively, of that of freshly isolated cells. On Day 6, the 7-ethoxyresorufin-O-deethylase and the aldrin epoxidase activities were still more than 50% that of freshly isolated hepatocytes. Exposure to phenobarbital on Days 3 to 6 increased the total cytochrome P-450 content twofold; exposure to 3-methylcholanthrene increased the activity of the corresponding cytochrome P-450 isoforms to 20 times that observed in untreated cultures and 6 times that observed in freshly isolated cells. Thus, given the ease with which they are prepared, the use of crude membrane fractions combined with culture medium supplemented with aprotinin and selenium can facilitate the preparation of reproducible cultures suitable for long-term in vitro pharmacotoxicologic studies using rat hepatocytes.</p>","PeriodicalId":77173,"journal":{"name":"In vitro cellular & developmental biology : journal of the Tissue Culture Association","volume":"29A 1","pages":"32-40"},"PeriodicalIF":0.0,"publicationDate":"1993-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02634369","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18683273","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fibroblast inhibition of tumor cells may be mediated by TGF-beta 1.","authors":"D Kirk,&nbsp;T Broberg,&nbsp;J C Irwin,&nbsp;W C Kenney,&nbsp;L W Jones","doi":"10.1007/BF02634364","DOIUrl":"https://doi.org/10.1007/BF02634364","url":null,"abstract":"","PeriodicalId":77173,"journal":{"name":"In vitro cellular & developmental biology : journal of the Tissue Culture Association","volume":"29A 1","pages":"4-6"},"PeriodicalIF":0.0,"publicationDate":"1993-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02634364","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19428997","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Differentiation of isolated murine embryonic palatal epithelium in culture: exogenous transforming growth factor alpha modulates matrix biosynthesis in defined experimental conditions.","authors":"M J Dixon,&nbsp;M J Carette,&nbsp;B B Moser,&nbsp;M W Ferguson","doi":"10.1007/BF02634371","DOIUrl":"https://doi.org/10.1007/BF02634371","url":null,"abstract":"<p><p>A novel culture technique, which supports the growth and differentiation of mouse embryonic palatal epithelial cells in the absence of either an extracellular matrix substratum or feeder layers, has been developed. Using this technique we have investigated the effects of exogenous transforming growth factor alpha (TGF alpha) and serum on extracellular matrix biosynthesis by primary cultures of mouse embryonic epithelial sheets under defined experimental conditions. In all culture treatments (chemically defined medium with and without TGF alpha or serum) the palatal epithelial sheets differentiated into three regionally distinct cell phenotypes after 36 h. Nasal and oral cells differentiated into pseudostratified, ciliated columnar, and stratified squamous keratinizing epithelium, respectively. In addition, basal medial edge epithelial (MEE) cells at the oral/nasal regional interface assumed an elongated cobblestoned phenotype. In serum-free medium, collagen types IV and V, laminin, fibronectin, and heparan sulphate proteoglycan were detected immunocytochemically throughout the entire epithelial sheet. Tenascin and collagen IX were present almost exclusively in MEE cells. Types I, II, and III collagen were completely absent. Addition of TGF alpha or serum universally increased the intensity of staining, most notably that for tenascin and collagen IX in MEE cells. These results indicate that mouse embryonic palatal epithelial sheets can be maintained under defined culture conditions during which they exhibit patterns of differentiation similar to those observed in vivo. TGF alpha, known to localize to the MEE in vivo, can modulate palatal extracellular matrix biosynthesis, particularly by the MEE, suggesting a regulatory role for this factor. The culture system is suitable for further investigating the effects of exogenous factors on mouse embryonic palatal epithelial cell bioactivity and differentiation.</p>","PeriodicalId":77173,"journal":{"name":"In vitro cellular & developmental biology : journal of the Tissue Culture Association","volume":"29A 1","pages":"51-61"},"PeriodicalIF":0.0,"publicationDate":"1993-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02634371","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19428998","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A superior method for single cell dispersal of rat and tumorous human anterior pituitary tissue.","authors":"S L Atkin,&nbsp;A M Landolt,&nbsp;R V Jeffreys,&nbsp;M C White","doi":"10.1007/BF02634363","DOIUrl":"https://doi.org/10.1007/BF02634363","url":null,"abstract":"","PeriodicalId":77173,"journal":{"name":"In vitro cellular & developmental biology : journal of the Tissue Culture Association","volume":"29A 1","pages":"1-3"},"PeriodicalIF":0.0,"publicationDate":"1993-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02634363","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19428994","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transforming growth factor-beta: signal transduction via protein kinase C in cultured embryonic vascular smooth muscle cells.","authors":"R W Wrenn,&nbsp;C L Raeuber,&nbsp;L E Herman,&nbsp;W J Walton,&nbsp;T H Rosenquist","doi":"10.1007/BF02634374","DOIUrl":"https://doi.org/10.1007/BF02634374","url":null,"abstract":"<p><p>Transforming growth factor-beta (TGF-beta), an ubiquitous regulatory peptide, has diverse effects on the differentiation and behavior of vascular smooth muscle cells (VSMC). However, the molecular mechanism through which TGF-alpha exerts its effects remains obscure. We investigated the phosphoinositide/protein kinase C [PKC] signaling pathway in the action of TGF-beta on cultured embryonic avian VSMC of differing lineage: a) thoracic aorta, derived from the neural crest; and b) abdominal aorta, derived from mesenchyme. The second messenger responsible for activation of PKC is sn-1,2-diacylglycerol [DAG]; TGF-beta increased the mass amounts of DAG in the membranes of neural crest-derived VSMC concurrent with translocation of PKC from the soluble to the membrane fraction, but TGF-beta had no effect on the DAG or PKC of mesenchyme-derived VSMC. TGF-beta potentiated the growth of platelet-derived growth factor (PDGF)-treated, neural crest-derived VSMC; but abolished PDGF-induced growth of mesenchymal cells. It is concluded that molecular and functional responses of VSMC to TGF-beta are heterogeneous and are functions of the embryonic lineage of the VSMC.</p>","PeriodicalId":77173,"journal":{"name":"In vitro cellular & developmental biology : journal of the Tissue Culture Association","volume":"29A 1","pages":"73-8"},"PeriodicalIF":0.0,"publicationDate":"1993-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02634374","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19429000","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cultures of bovine tracheal epithelium with differentiated ultrastructure and ion transport.","authors":"M Kondo,&nbsp;W E Finkbeiner,&nbsp;J H Widdicombe","doi":"10.1007/BF02634367","DOIUrl":"https://doi.org/10.1007/BF02634367","url":null,"abstract":"<p><p>Tracheal epithelial cells were grown on Nuclepore filters coated with human placental collagen. When grown immersed in medium containing fetal bovine serum, cells displayed an undifferentiated ultrastructure (no cilia and a cell height of approximately 10 microns). Short-circuit current (Isc) was approximately 1/10 that of the native epithelium. By contrast, when grown in hormonally defined, serum-free medium with an air interface, cells showed Isc equal to or greater than the original tissue, possessed cilia, and had a cell height of approximately 50 microns. Responses in Isc to mediators were similar to those of the original tissue, but differed from those of dog or human tracheal epithelium. Given the ready availability and low cost of the native tissues, bovine tracheal cultures grown in serum-free medium with an air interface should prove useful in studies of airway epithelial physiology.</p>","PeriodicalId":77173,"journal":{"name":"In vitro cellular & developmental biology : journal of the Tissue Culture Association","volume":"29A 1","pages":"19-24"},"PeriodicalIF":0.0,"publicationDate":"1993-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02634367","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19428995","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Organ-specific change in Dolichos biflorus lectin binding by myocardial endothelial cells during in vitro cultivation.","authors":"J Plendl,&nbsp;L Hartwell,&nbsp;R Auerbach","doi":"10.1007/BF02634368","DOIUrl":"https://doi.org/10.1007/BF02634368","url":null,"abstract":"<p><p>Endothelial cells of the NMRI mouse strain express a cell surface glycoprotein recognized by the lectin Dolichos biflorus agglutinin (DBA). This study documents a marked organ-specific increase in DBA-specific lectin binding of myocardium-derived endothelial cells (MEC) of the NMRI/GSF mouse during in vitro cultivation. An up to 20-fold increase in DBA binding sites is observed in long-term culture, an increase not found in other NMRI-derived endothelial cell lines (e.g., brain, aorta). The increase appears restricted to DBA in that binding with other lectins (PNA, WGA) was unaltered. NMRI MEC cultures maintain typical endothelial cell attributes such as cobblestone morphology on confluence, expression of endothelial cell-specific surface markers, and production of angiotensin-converting enzyme. Cultures routinely become aneuploid within 4 passages, several passages before upregulation of the DBA binding site(s). Myocardial endothelial cells sorted to obtain DBAhi and DBAlo cell populations generally maintained their sorted phenotype for 3 to 4 passages. Limiting dilution cloning resulted in clones varying in DBA expression. Clones for DBAhi expression maintained their DBA affinity for at least 10 passages (> 30 doublings), whereas DBAlo clones gave rise to varying numbers of DBAhi cells within 2 to 4 passages. We hypothesize that the change in DBA affinity accompanies in vitro aging, that the change is independent of alterations in karyotype, and that the increase in DBA affinity may reflect a change in one or more other endothelial cell properties. Additional studies will be necessary to determine whether the in vitro changes are correlated with specific functional alterations and whether they accurately reflect progressive changes of MEC in vivo.</p>","PeriodicalId":77173,"journal":{"name":"In vitro cellular & developmental biology : journal of the Tissue Culture Association","volume":"29A 1","pages":"25-31"},"PeriodicalIF":0.0,"publicationDate":"1993-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02634368","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19428996","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Clonogenic assay of leukemic cells in serum-free culture.","authors":"Y C Dai,&nbsp;X Q Lu,&nbsp;Z Fang","doi":"10.1007/BF02631048","DOIUrl":"https://doi.org/10.1007/BF02631048","url":null,"abstract":"","PeriodicalId":77173,"journal":{"name":"In vitro cellular & developmental biology : journal of the Tissue Culture Association","volume":"28A 11-12","pages":"685-6"},"PeriodicalIF":0.0,"publicationDate":"1992-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02631048","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12653886","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Skin histoculture assay for studying the hair cycle.","authors":"L Li,&nbsp;R Paus,&nbsp;A Slominski,&nbsp;R M Hoffman","doi":"10.1007/BF02631052","DOIUrl":"https://doi.org/10.1007/BF02631052","url":null,"abstract":"","PeriodicalId":77173,"journal":{"name":"In vitro cellular & developmental biology : journal of the Tissue Culture Association","volume":"28A 11-12","pages":"695-8"},"PeriodicalIF":0.0,"publicationDate":"1992-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02631052","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12653893","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of normal breast epithelial cells in primary cultures: differentiation and growth factor receptors studies.","authors":"P Berthon,&nbsp;G Pancino,&nbsp;P de Cremoux,&nbsp;A Roseto,&nbsp;C Gespach,&nbsp;F Calvo","doi":"10.1007/BF02631059","DOIUrl":"https://doi.org/10.1007/BF02631059","url":null,"abstract":"<p><p>The growth and differentiation of normal human mammary epithelial cells (HMEC) were studied after propagation of serial cultures from breast tissue biopsies from 42 mammoplasty patients. Cells were grown for up to 7 mo. in low calcium medium. HMEC cultures displayed heterogeneous growth patterns, according to the average doubling time of 44 +/- 6 h for 32 generations. Proliferation peaked at Day 30. HMEC maintained a normal karyotype and were organized in ductlike structures when cultured in collagen gel matrix. The cultures retained several phenotype traits of the epithelial lineage, including the expression of cytokeratins 18 and 19, specific mammary gland antigens, as shown by indirect HMEC immunostaining by the monoclonal antibodies DF3, EMA, 7B10, and 1BE12. Estrogen receptors were undetectable, whereas progesterone receptors were present at very low density. High-affinity cell surface receptors for epidermal growth factor (EGF) (Kd = 1.1 x 10(-10) M) were observed at a density of 50,000 to 100,000 sites per cell. Accordingly, [3H]thymidine incorporation in HMEC was optimally stimulated by EGF at concentrations of 10(-11) to 10(-10) M. HMEC were also seen to possess functional VIP receptors linked to the adenylate cyclase system, as we previously observed in seven human breast cancer cell lines. These results show that long-term cultures of HMEC provide useful models for studying the growth and differentiation of the normal human mammary gland, and the role of growth factors and hormones in these functions.</p>","PeriodicalId":77173,"journal":{"name":"In vitro cellular & developmental biology : journal of the Tissue Culture Association","volume":"28A 11-12","pages":"716-24"},"PeriodicalIF":0.0,"publicationDate":"1992-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02631059","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12457557","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}