Name: stabilizing hepatocellular phenotype using optimized synthetic surfaces
Uploaded: 2014-09-26T14:00:00
Description: Watch this Scientific Journal Video about Stabilizing Hepatocellular Phenotype Using Optimized Synthetic Surfaces at JoVE.com

DCH, MB og FK ble støttet av en EPSRC Følg på fondet. BL-V og DS ble hver støttet av MRC Doktorgradsstipend. KC ble støttet av midler fra Storbritannia regenerativ medisin Platform.

1. Syntese av PHNGAD (Poly [1,6-hexanodiol / neopentylglykol / di (etylenglykol) -alt-adipinsyre] diol) <img alt="Skjema 1" fo:content-width="5in" src="/files/ftp_upload/51723/51723scheme1.jpg" width="500" /> Skjema 1: Syntese av PHNAGD Skjematisk representasjon av syntesen av PHNAGD.. PHNAGD ble fremstilt ved omsetning av 1,6-Hexanodiol, dietylenglykol, neoppentyl glykol og adipinsyre. PHNAGD, Poly [1,6-hexanodiol / neopentylglykol / di (etylenglykol) -alt-adipinsyre] diol....

<ol>
	<li>Zhou, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SUMOylation+of+HNF4&#945;+regulates+protein+stability+and+hepatocyte+function.">SUMOylation of HNF4&#945; regulates protein stability and hepatocyte function.</a> J Cell Sci. 125 (15), 3630-3635 (2012).</li><li>Banerjee, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+influence+of+hydrogel+modulus+on+the+proliferation+and+differentiation+of+encapsulated+neural+stem+cells.">The influence of hydrogel modulus on the proliferation and differentiation of encapsulated neural stem cells.</a> Biomaterials. 30 (27), 4695-4699 (2009).</li><li>Shanbhag, M. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+Progenitor+Cells+Grown+on+Hydrogel+Surfaces+Respond+to+the+Product+of+the+Transgene+of+Encapsulated+Genetically+Engineered+Fibroblasts.">Neural Progenitor Cells Grown on Hydrogel Surfaces Respond to the Product of the Transgene of Encapsulated Genetically Engineered Fibroblasts.</a> Biomacromolecules. 11 (11), 2936-2943 (2010).</li><li>Battista, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+matrix+composition+of+3D+constructs+on+embryonic+stem+cell+differentiation.">The effect of matrix composition of 3D constructs on embryonic stem cell differentiation.</a> Biomaterials. 26 (31), 6194-6207 (2005).</li><li>Tian, W. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyaluronic+acid+hydrogel+as+Nogo-66+receptor+antibody+delivery+system+for+the+repairing+of+injured+rat+brain:+in+vitro.">Hyaluronic acid hydrogel as Nogo-66 receptor antibody delivery system for the repairing of injured rat brain: in vitro.</a> Journal of Controlled Release. 102 (1), 13-22 (2005).</li><li>Keshaw, H., Forbes, A., Day, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Release+of+angiogenic+growth+factors+from+cells+encapsulated+in+alginate+beads+with+bioactive+glass.">Release of angiogenic growth factors from cells encapsulated in alginate beads with bioactive glass.</a> Biomaterials. 26 (19), 4171-4179 (2005).</li><li>Baharvand, H., Hashemi, S. M., Kazemi Ashtiani, S., Farrokhi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differentiation+of+human+embryonic+stem+cells+into+hepatocytes+in+2D+and+3D+culture+systems+in+vitro.">Differentiation of human embryonic stem cells into hepatocytes in 2D and 3D culture systems in vitro.</a> The International Journal of Developmental Biology. 50 (7), 645-652 (2006).</li><li>Cameron, K., Travers, P., Chander, C., Buckland, T., Campion, C., Noble, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Directed+osteogenic+differentiation+of+human+mesenchymal+stem/precursor+cells+on+silicate+substituted+calcium+phosphate.">Directed osteogenic differentiation of human mesenchymal stem/precursor cells on silicate substituted calcium phosphate.</a> Journal of Biomedical Materials Research Part A. 101 (1), 13-22 (2013).</li><li>Pernagallo, S., Unciti-Broceta, A., Diaz-Mochon, J. J., Bradley, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deciphering+cellular+morphology+and+biocompatibility+using+polymer+microarrays.">Deciphering cellular morphology and biocompatibility using polymer microarrays.</a> Biomedical Materials. 3 (3), 034112 (2008).</li><li>Li, Z., Guo, X., Matsushita, S., Guan, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differentiation+of+cardiosphere-derived+cells+into+a+mature+cardiac+lineage+using+biodegradable+poly(N-isopropylacrylamide)+hydrogels.">Differentiation of cardiosphere-derived cells into a mature cardiac lineage using biodegradable poly(N-isopropylacrylamide) hydrogels.</a> Biomaterials. 32 (12), 3220-3232 (2011).</li><li>Tare, R. S., Khan, F., Tourniaire, G., Morgan, S. M., Bradley, M., Oreffo, R. O. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+microarray+approach+to+the+identification+of+polyurethanes+for+the+isolation+of+human+skeletal+progenitor+cells+and+augmentation+of+skeletal+cell+growth.">A microarray approach to the identification of polyurethanes for the isolation of human skeletal progenitor cells and augmentation of skeletal cell growth.</a> Biomaterials. 30 (6), 1045-1055 (2009).</li><li>Khan, F., Tare, R. S., Kanczler, J. M., Oreffo, R. O. C., Bradley, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strategies+for+cell+manipulation+and+skeletal+tissue+engineering+using+high-throughput+polymer+blend+formulation+and+microarray+techniques.">Strategies for cell manipulation and skeletal tissue engineering using high-throughput polymer blend formulation and microarray techniques.</a> Biomaterials. 31 (8), 2216-2228 (2010).</li><li>Zhang, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+thermoresponsive+and+chemically+defined+hydrogel+for+long-term+culture+of+human+embryonic+stem+cells.">A thermoresponsive and chemically defined hydrogel for long-term culture of human embryonic stem cells.</a> Nature Communications. 4 (1335), (2013).</li><li>Medine, C. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developing+high-fidelity+hepatotoxicity+models+from+pluripotent+stem+cells.">Developing high-fidelity hepatotoxicity models from pluripotent stem cells.</a> Stem Cells Translational Medicine. 2 (7), 505-509 (2013).</li><li>Hay, D. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unbiased+screening+of+polymer+libraries+to+define+novel+substrates+for+functional+hepatocytes+with+inducible+drug+metabolism.">Unbiased screening of polymer libraries to define novel substrates for functional hepatocytes with inducible drug metabolism.</a> Stem Cell Research. 6 (2), 92-102 (2011).</li><li>Lucendo-Villarin, B., Khan, F., Pernagallo, S., Bradley, M., Iredale, J. P., Hay, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maintaining+hepatic+stem+cell+gene+expression+on+biological+and+synthetic+substrata.">Maintaining hepatic stem cell gene expression on biological and synthetic substrata.</a> BioResearch Open Access. 1 (1), 50-53 (2012).</li><li>Hay, D. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Highly+efficient+differentiation+of+hESCs+to+functional+hepatic+endoderm+requires+ActivinA+and+Wnt3a+signaling.">Highly efficient differentiation of hESCs to functional hepatic endoderm requires ActivinA and Wnt3a signaling.</a> Proceedings of the National Academy of Sciences. 105 (34), 12301-12306 (2008).</li><li>Szkolnicka, D., Zhou, W., Lucendo-Villarin, B., Hay, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pluripotent+Stem+Cell&#8211;Derived+Hepatocytes:+Potential+and+Challenges+in+Pharmacology.">Pluripotent Stem Cell&#8211;Derived Hepatocytes: Potential and Challenges in Pharmacology.</a> Annu Rev Pharmecol Toxicol. 53, 147-149 (2013).</li><li>Szkolnicka, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accurate+prediction+of+drug-induced+liver+injury+using+stem+cell-derived+populations.">Accurate prediction of drug-induced liver injury using stem cell-derived populations.</a> Stem Cells Translational Medicine. 3 (2), 141-148 (2014).</li><li>Medine, C. N., Lucendo-Villarin, B., Zhou, W., West, C. C., Hay, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Robust+Generation+of+Hepatocyte-like+Cells+from+Human+Embryonic+Stem+Cell+Populations.">Robust Generation of Hepatocyte-like Cells from Human Embryonic Stem Cell Populations.</a> Journal of Visualized Experiments. (56), (2011).</li><li>Freed, L. E., Vunjak-Novakovic, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Culture+of+organized+cell+communities.">Culture of organized cell communities.</a> Advanced Drug Delivery Reviews. 33 (1-2), 15-30 (1998).</li><li>Braam, S. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recombinant+Vitronectin+Is+a+Functionally+Defined+Substrate+That+Supports+Human+Embryonic+Stem+Cell+Self-Renewal+via+&#945;V&#946;5+Integrin.">Recombinant Vitronectin Is a Functionally Defined Substrate That Supports Human Embryonic Stem Cell Self-Renewal via &#945;V&#946;5 Integrin.</a> Stem Cells. 26 (9), 2257-2265 (2008).</li><li>Rodin, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clonal+culturing+of+human+embryonic+stem+cells+on+laminin-521/E-cadherin+matrix+in+defined+and+xeno-free+environment.">Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment.</a> Nature Communications. 5 (3195), (2014).</li><li>Thaburet, J. -. F. O., Mizomoto, H., Bradley, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-Throughput+Evaluation+of+the+Wettability+of+Polymer+Libraries.">High-Throughput Evaluation of the Wettability of Polymer Libraries.</a> Macromolecular Rapid Communication. 25 (1), 336-370 (2003).</li><li>Lim, J. Y., Donahue, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+Sensing+and+Response+to+Micro-+and+Nanostructured+Surfaces+Produced+by+Chemical+and+Topographic+Patterning.">Cell Sensing and Response to Micro- and Nanostructured Surfaces Produced by Chemical and Topographic Patterning.</a> Tissue Engineering. 13 (8), 1879-1891 (2007).</li><li>Teixeira, A. I., Abrams, G. A., Bertics, P. J., Murphy, C. J., Nealey, P. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epithelial+contact+guidance+on+well-defined+micro-+and+nanostructured+substrates.">Epithelial contact guidance on well-defined micro- and nanostructured substrates.</a> Journal of Cell Science. 116 (10), 1881-1892 (2003).</li><li>Biggs, M. J. P., Richards, R. G., Wilkinson, C. D. W., Dalby, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Focal+adhesion+interactions+with+topographical+structures:+a+novel+method+for+immuno-SEM+labelling+of+focal+adhesions+in+S-phase+cells.">Focal adhesion interactions with topographical structures: a novel method for immuno-SEM labelling of focal adhesions in S-phase cells.</a> Journal of Microscopy. 231 (1), 28-37 (2008).</li><li>Karuri, N. W., Porri, T. J., Albrecht, R. M., Murphy, C. J., Nealey, P. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nano-+and+microscale+holes+modulate+cell-substrate+adhesion,+cytoskeletal+organization,+and+-beta1+integrin+localization+in+SV40+human+corneal+epithelial+cells.">Nano- and microscale holes modulate cell-substrate adhesion, cytoskeletal organization, and -beta1 integrin localization in SV40 human corneal epithelial cells.</a> IEEE Transactions on Nanobioscience. 5 (4), 273-280 (2006).</li><li>Hamilton, D. W., Brunette, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+substratum+topography+on+osteoblast+adhesion+mediated+signal+transduction+and+phosphorylation.">The effect of substratum topography on osteoblast adhesion mediated signal transduction and phosphorylation.</a> Biomaterials. 28 (1), 1806-1819 (2007).</li><li>Engler, A. J., Sen, S., Sweeney, H. L., Discher, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Matrix+elasticity+directs+stem+cell+lineage+specification.">Matrix elasticity directs stem cell lineage specification.</a> Cell. 126 (4), 677-689 (2006).</li><li>Dang, J. M., Leong, K. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myogenic+Induction+of+Aligned+Mesenchymal+Stem+Cell+Sheets+by+Culture+on+Thermally+Responsive+Electrospun+Nanofibers.">Myogenic Induction of Aligned Mesenchymal Stem Cell Sheets by Culture on Thermally Responsive Electrospun Nanofibers.</a> Advanced Materials. 19 (19), 2775-2779 (2007).</li><li>Azevedo, E. C., Nascimento, E. M., Chierice, G. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=UV+and+gamma+irradiation+effects+on+surface+properties+of+polyurethane+derivate+from+castor+oil.">UV and gamma irradiation effects on surface properties of polyurethane derivate from castor oil.</a> Pol&#237;meros. 23 (3), 305-311 (2013).</li><li>Rosu, L., Cascaval, C. N., Ciobanu, C., Rosu, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+UV+radiation+on+the+semi-interpenetrating+polymer+networks+based+on+polyurethane+and+epoxy+maleate+of+bisphenol+A.">Effect of UV radiation on the semi-interpenetrating polymer networks based on polyurethane and epoxy maleate of bisphenol A.</a> Journal of Photochemistry and Photobilogy A: Chemistry. 169 (2), 177-185 (2005).</li><li>Yang, X. F., Tallman, D. E., Bierwagen, G. P., Croll, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blistering+and+degradation+of+polyurethane+coatings+under+different+accelerated+weathering+tests.">Blistering and degradation of polyurethane coatings under different accelerated weathering tests.</a> Polymer Degradation and Stability. 77 (1), 103-109 (2002).</li></ol>

Mange av dagens metoder som brukes til å generere hepatocytter fra stamceller stole på udefinerte matriser av animalsk opprinnelse. Slike underlag kan være kostbart og svært variabel, påvirker cellefunksjon og stabilitet, som representerer en betydelig barriere for søknaden. Derfor gjennomførte vi en skjerm for syntetiske materialer som støtter kulturen i stamcelle avledet hepatocytter. Vi har identifisert en enkel polyuretan (PU134), dannet ved polymerisering av PHNGAD, MDI, og et drøyemiddel, som i kombinasjo...

Biologisk materiale har blitt mye brukt i vedlikehold og differensiering av pluripotente stamceller en. Mens slik at disse biologiske substrater ofte inneholder en myriade av udefinerte komponenter. Matrigel er et vanlig substrat for stamcellekultur og differensiering. Dessverre, det er variabel sammensetning påvirker cellefunksjon og fenotype. Selv om en rekke av alternative og mer definert biologiske matriser er blitt brukt 2-7, gjør deres animalsk opprinnelse eller dårlig skalerbarhet dem uegnede kandidater for industriell produksjon. Derfor identifisering av syntetiske alternativer, med definert sammensetning og pålitelig ytelse, er sentrale mål i stamcelleforskning. I et forsøk på å overvinne begrensninger av udefinerte cellekultur substrater, har tverr samarbeid mellom kjemi og biologi identifiserte syntetiske materialer med evne til å støtte celle fenotype. Synthetic underlag er skalerbar, kostnadseffektiv, og kan produseres til komplekse 3D-strukturer, etterligne in vivo miljøet. På grunn av disse egenskapene syntetiske substrater er blitt mye brukt for å støtte og drive differensiering av mange celletyper, 8-10. Avanserte og høy gjennomstrømning analyser har tilrettelagt rask screening av syntetiske materialer, fra store biblioteker, og levert nye materialer med fleksible eiendommer med brede programmer i biomedisinsk forskning og utvikling 11-13. Utnytte høy gjennomstrømning, polymer microarray screening teknologi, vi raskt identifisert en enkel polyuretan (PU134), egnet for vedlikehold av menneskelige stamceller avledet hepatocytter. Denne polymer ble funnet å være overlegne i forhold til dyre avledet substrater med hensyn til hepatocytter differensiering og funksjon 14-16. Vi har senere optimalisert belegg forhold, topografi og steriliseringsprosess for å få tilgang effekterpå polymer ytelse i å stabilisere hepatocytter funksjon og levetid. Dette har betydelige konsekvenser med hensyn til å forstå grunnleggende av hepatocytter biologi for cellebasert modellering og regenerativ medisin applikasjoner. Teknologien som er beskrevet her, representerer et eksempel på hvordan overflaten av en syntetisk polymer kan være optimalisert for å bevare celle fenotype. Vi tror at kombinasjonen av denne teknologien med en effektiv serumfritt hepatocytter differensiering protokollen har potensial til å gi en skalerbar produksjon av hepatocytter for anvendelse i in vitro-modeller og regenerativ medisin.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Synthesis, preparation, coating and characterization of polymer PU134 coated coverslips</td>
		</tr>
		<tr>
			<td>Shaker</td>
			<td>Edmun B&#252;hler</td>
			<td>KS-15</td>
			<td></td>
		</tr>
		<tr>
			<td>Irradiator</td>
			<td>CIS Biointernational</td>
			<td>IBL 637&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Spin coater</td>
			<td>Specialty Coating System</td>
			<td>&#160;P-6708</td>
			<td></td>
		</tr>
		<tr>
			<td>Scanning Electron Microscope&#160;</td>
			<td>Philips</td>
			<td>XL30CPSEM</td>
			<td></td>
		</tr>
		<tr>
			<td>Atomic Force Microscope</td>
			<td>DimensionV Nanoscope, VEECO</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>p4-GLO CYP3A4</td>
			<td>Promega</td>
			<td>V8902</td>
			<td></td>
		</tr>
		<tr>
			<td>UV bulb</td>
			<td>ESCO</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>NanoScope analysis software</td>
			<td>VEECO</td>
			<td></td>
			<td>version 1.20</td>
		</tr>
		<tr>
			<td>Fluorescence microscope</td>
			<td>Olympus</td>
			<td>TH45200</td>
			<td>Use Volocity 4 Software</td>
		</tr>
		<tr>
			<td>Tissue culture plates</td>
			<td>Corning, UK&#160;</td>
			<td>3527</td>
			<td></td>
		</tr>
		<tr>
			<td>glass slides</td>
			<td>Scientific Laboratory Supplies</td>
			<td>MIC3308</td>
			<td></td>
		</tr>
		<tr>
			<td>Diethylene glycol</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>93171</td>
			<td></td>
		</tr>
		<tr>
			<td>&#160;1,6-hexanediol</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>240117</td>
			<td></td>
		</tr>
		<tr>
			<td>Neopentyl glycol</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>408255</td>
			<td></td>
		</tr>
		<tr>
			<td>Adipic acid</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>9582</td>
			<td></td>
		</tr>
		<tr>
			<td>anhydrous N,N-Dimethylformamide</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>227056</td>
			<td></td>
		</tr>
		<tr>
			<td>Diethyl ether</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>676845</td>
			<td></td>
		</tr>
		<tr>
			<td>titanium (IV) butoxide&#160;</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>244112</td>
			<td></td>
		</tr>
		<tr>
			<td>1,4-butanediol&#160;</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>493732</td>
			<td></td>
		</tr>
		<tr>
			<td>Vacuum oven</td>
			<td>Thermoscientific</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>4,4&#8217;-Methylenebis(phenyl isocyanate)</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>101688</td>
			<td></td>
		</tr>
		<tr>
			<td>Tetrahydrofurane</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>401757</td>
			<td></td>
		</tr>
		<tr>
			<td>Sputter coater</td>
			<td>Bal-Tec SCD 050</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Inmunostaining</td>
		</tr>
		<tr>
			<td>Phosphate buffer saline (-MgCl2, -CaCl2)</td>
			<td>Gibco</td>
			<td>10010031</td>
			<td>&#160;Store at room temperature</td>
		</tr>
		<tr>
			<td>PBST, PBS made up with 0.1% TWEEN 20&#160;</td>
			<td>&#160; Scientific Laboratory Supplies Ltd</td>
			<td>EC607&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>&#160; Scientific Laboratory Supplies Ltd</td>
			<td>CHE5010</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine Serum Albumin</td>
			<td>Sigma-Aldrich, UK</td>
			<td>A7906</td>
			<td></td>
		</tr>
		<tr>
			<td>MOWIOL 488 DAPI</td>
			<td>Calbiochem</td>
			<td>475904</td>
			<td>Made up in Tris HCL and glycerol as per manufacturers instructions</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Cell culture and Functional assay</td>
		</tr>
		<tr>
			<td>CYP3A activity pGLO kit</td>
			<td>Promega</td>
			<td>V8902</td>
			<td></td>
		</tr>
		<tr>
			<td>Hepatozyme</td>
			<td>Gibco</td>
			<td>17705021</td>
			<td></td>
		</tr>
		<tr>
			<td>TryLE express</td>
			<td>Life Technologies</td>
			<td><a href="http://www.lifetechnologies.com/order/catalog/product/12604013">12604013</a></td>
			<td></td>
		</tr>
	</tbody>
</table>

stabilizing hepatocellular phenotype using optimized synthetic surfaces

Currently, one of the major limitations in cell biology is maintaining differentiated cell phenotype. Biological matrices are commonly used for culturing and maintaining primary and pluripotent stem cell derived hepatocytes. While biological matrices are useful, they permit short term culture of hepatocytes, limiting their widespread application. We have attempted to overcome the limitations using a synthetic polymer coating. Polymers represent one of the broadest classes of biomaterials and possess a wide range of mechanical, physical and chemical properties, which can be fine-tuned for purpose. Importantly, such materials can be scaled to quality assured standards and display batch-to-batch consistency. This is essential if cells are to be expanded for high through-put screening in the pharmaceutical testing industry or for cellular based therapy. Polyurethanes (PUs) are one group of materials that have shown promise in cell culture. Our recent progress in optimizing a polyurethane coated surface, for long-term culture of human hepatocytes displaying stable phenotype, is presented and discussed.

Polymerløsningsmidlet påvirker topografien av polymeren belagte overflaten Polyuretan 134 ble oppløseliggjort i kloroform, enten alene eller i kombinasjon med toluen eller tetrahydrofuran eller diklormetan, og de glassplater spin-belagt med de forskjellige formuleringer. Scanning elektronmikroskopi (SEM) og atomic force microscopy (AFM) ble brukt for å karakterisere de fysikalske egenskaper for de polymere belegg (figur 1). Den oppnådde ved å bruke tol...

Watch this Scientific Journal Video about Stabilizing Hepatocellular Phenotype Using Optimized Synthetic Surfaces at JoVE.com

Stabilizing Hepatocellular Phenotype Using Optimized Synthetic Surfaces

Denne artikkelen vil fokusere på å utvikle polymer belagt overflater for langsiktig, stabil kultur for stamcelle avledet humane hepatocytter.

Stabiliserende Hepatocellulært Fenotype Ved hjelp av optimalisert Syntetiske Overflater

Currently, one of the major limitations in cell biology is maintaining differentiated cell phenotype. Biological matrices are commonly used for culturing ...

Research

JoVE Journal

Chemistry

Covalent Binding of BMP-2 on Surfaces Using a Self-assembled Monolayer Approach

Covalent Binding av BMP-2 på overflater Bruke en Self-montert med ett lag Approach

Bone morphogenetic protein 2 (BMP-2) is a growth factor embedded in the extracellular matrix of bone tissue. BMP-2 acts as trigger of mesenchymal cell ...

Towards Biomimicking Wood: Fabricated Free-standing Films of Nanocellulose, Lignin, and a Synthetic Polycation

Mot Biomimicking Wood: Fabricated Frittstående Films av nanocellulose, lignin, og en syntetisk polykation

Woody materials are comprised of plant cell walls that contain a layered secondary cell wall composed of structural polymers of polysaccharides and ...

In Situ SIMS and IR Spectroscopy of Well-defined Surfaces Prepared by Soft Landing of Mass-selected Ions

In Situ SIMS and IR Spectroscopy of Well-defined Surfaces Prepared by Soft Landing of Mass-selected Ions

In situ SIMS og IR spektroskopi av godt definerte overflater som er Soft Landing av Mass-utvalgte ioner

Soft landing of mass-selected ions onto surfaces is a powerful approach for the highly-controlled preparation of materials that are inaccessible using ...

Synthetic Methodology for Asymmetric Ferrocene Derived Bio-conjugate Systems via Solid Phase Resin-based Methodology

Syntetisk Methodology for Asymmetric Ferrocene Avledet Bio-konjugerte systemer via Solid Phase Resin-basert metode

Early detection is a key to successful treatment of most diseases, and is particularly imperative for the diagnosis and treatment of many types of cancer. ...

Optimization of Synthetic Proteins: Identification of Interpositional Dependencies Indicating Structurally and/or Functionally Linked Residues

Optimalisering av Syntetisk Proteiner: Identifisering av Interpositional Avhengig Hente Strukturelt og / eller Funksjonelt Koblede Rester

Protein alignments are commonly used to evaluate the similarity of protein residues, and the derived consensus sequence used for identifying functional ...

An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group

An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group

En optimalisert Protokoll for effektiv Radiomerking av gull nanopartikler ved hjelp av en<sup&gt; 125</sup&gt; I-merket Azid Prosthetic Gruppe

Here, we demonstrate a detailed protocol for the radiosynthesis of a 125I-labeled azide prosthetic group and its application to the efficient ...

Color Spot Test As a Presumptive Tool for the Rapid Detection of Synthetic Cathinones

Farge stikkprøve som Presumptive verktøy for rask påvisning av syntetisk katinoner

Synthetic cathinones are a large class of new psychoactive substances (NPS) that are increasingly prevalent in drug seizures made by law enforcement and ...

Characterization of Synthetic Polymers via Matrix Assisted Laser Desorption Ionization Time of Flight (MALDI-TOF) Mass Spectrometry

Characterization of Synthetic Polymers via Matrix Assisted Laser Desorption Ionization Time of Flight MALDI-TOF Mass Spectrometry

Karakteristikk av syntetiske polymerer via Matrix assistert Laser desorpsjon Ionization tiden av fly (MALDI-TOF) massespektrometri

There are many techniques that can be employed in the characterization of synthetic homopolymers, but few provide as useful of information for end group ...

Light-driven Molecular Motors on Surfaces for Single Molecular Imaging

Lys-drevet molekylær motorer på overflater for enkelt molekylær Imaging

The design and synthesis of a synthetic system that aims for the direct visualization of a synthetic rotary motor at the single molecule level on surfaces ...

A Facile Synthetic Method to Obtain Bismuth Oxyiodide Microspheres Highly Functional for the Photocatalytic Processes of Water Depuration

Lettvinte syntetiske enmetoden å få Vismut Oxyiodide mikrosfærer svært funksjonell for Photocatalytic prosesser av vann rensingen

Bismuth oxyhalide (BiOI) is a promising material for sunlight-driven-environmental photocatalysis. Given that the physical structure of this kind of ...