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Publication

Consensus guidelines for the use and interpretation of angiogenesis assays

Journal: Angiogenesis
Publication Date: 15 May 2018
Author: Nowak-Sliwinska P , et al.

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Authors	Nowak-Sliwinska P, Alitalo K, Allen E, Anisimov A, Aplin AC, Auerbach R, Augustin HG, Bates DO, van Beijnum JR, Bender RHF, Bergers G, Bikfalvi A, Bischoff J, Böck BC, Brooks PC, Bussolino F, Cakir B, Carmeliet P, Castranova D, Cimpean AM, Cleaver O, Coukos G, Davis GE, De Palma M, Dimberg A, Dings RPM, Djonov V, Dudley AC, Dufton NP, Fendt SM, Ferrara N, Fruttiger M, Fukumura D, Ghesquière B, Gong Y, Griffin RJ, Harris AL, Hughes CCW, Hultgren NW, Iruela-Arispe ML, Irving M, Jain RK, Kalluri R, Kalucka J, Kerbel RS, Kitajewski J, Klaassen I, Kleinmann HK, Koolwijk P, Kuczynski E, Kwak BR, Marien K, Melero-Martin JM, Munn LL, Nicosia RF, Noel A, Nurro J, Olsson AK, Petrova TV, Pietras K, Pili R, Pollard JW, Post MJ, Quax PHA, Rabinovich GA, Raica M, Randi AM, Ribatti D, Ruegg C, Schlingemann RO, Schulte-Merker S, Smith LEH, Song JW, Stacker SA, Stalin J, Stratman AN, Van de Velde M, van Hinsbergh VWM, Vermeulen PB, Waltenberger J, Weinstein BM, Xin H, Yetkin-Arik B, Yla-Herttuala S, Yoder MC, Griffioen AW
Journal	Angiogenesis
Publication Date	15 May 2018
Research Area(s) include	Cell Physiology Research, Review Papers, Techniques and Methods
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XF 96 well (XF Pro/XFe96/XF96), Seahorse XFe24/XF24, Seahorse XFp
Mode	N/A
Cell Line(s) include	N/A (review)
Cell Type(s) include	N/A (review)
Species	N/A (review)
Assay / Method	Cell Mitochondrial Stress Test, Glycolysis Stress Test
Cell Seeding Density	3.0x10^4 cells/well
Plate Coating	Not Specified

Authors	L H Lehmann, Z H Jebessa, M M Kreusser, A Horsch, T He, M Kronlage, M Dewenter, V Sramek, U Oehl, J Krebs-Haupenthal, A H von der Lieth, A Schmidt, Q Sun, J Ritterhoff, D Finke, M Volkers, A Jungmann, S W Sauer, C Thiel, A Nickel, M Kohlhaas, M Schafer, C Sticht, C Maack, N Gretz, M Wagner, A El-Armouche, L S Maier, J E C Londono, B Meder, M Freichel, H J Grone, P Most, O J Muller, S Herzig, E E M Furlong, H A Katus, and J Backs
Journal	Nat Med
Publication Date	04 Dec 2017
Research Area(s) include	Cancer Research, Translational Research
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XFe24/XF24
Cell Line(s) include	Neonatal rat ventricular myocytes (NRVM)
Cell Type(s) include	Cardiac cells
Species	Rat
Assay / Method	Cell Mitochondrial Stress Test
Cell Seeding Density	3.0x10^4 cells/well
Plate Coating	Not Specified

Authors	L Barbier-Torres, P Iruzubieta, D Fernandez-Ramos, T C Delgado, D Taibo, V Guitierrez-de-Juan, M Varela-Rey, M Azkargorta, N Navasa, P Fernandez-Tussy, I Zubiete-Franco, J Simon, F Lopitz-Otsoa, S Lachiondo-Ortega, J Crespo, S Masson, M V McCain, E Villa, H Reeves, F Elortza, M I Lucena, M I Hernandez-Alvarez, A Zorzano, R J, andrade, S C Lu, J M Mato, J Anguita, M Rincon, and M L Martinez-Chantar
Journal	Nat Commun
Publication Date	12 Dec 2017
Research Area(s) include	Cell Physiology Research, Drug Discovery, Toxicology Research
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XFe24/XF24
Cell Line(s) include	Hepatocytes
Cell Type(s) include	Hepatocytes
Species	Mouse
Assay / Method	Cell Mitochondrial Stress Test
Cell Seeding Density	2.0x10^4 cells/well
Plate Coating	Collagen

Authors	Codo AC, Davanzo GG, Monteiro LB, de Souza GF, Muraro SP, Virgilio-da-Silva JV, Prodonoff JS, Carregari VC, de Biagi Junior CAO, Crunfli F, Jimenez Restrepo JL, Vendramini PH, Reis-de-Oliveira G, Bispo Dos Santos K, Toledo-Teixeira DA, Parise PL, Martini MC, Marques RE, Carmo HR, Borin A, Coimbra LD, Boldrini VO, Brunetti NS, Vieira AS, Mansour E, Ulaf RG, Bernardes AF, Nunes TA, Ribeiro LC, Palma AC, Agrela MV, Moretti ML, Sposito AC, Pereira FB, Velloso LA, Vinolo MAR, Damasio A, Proenca-Modena JL, Carvalho RF, Mori MA, Martins-de-Souza D, Nakaya HI, Farias AS and Moraes-Vieira PM
Journal	Cell Metab
Publication Date	01 Sep 2020
Research Area(s) include	Infectious Disease Research
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XFe24/XF24
Cell Line(s) include	Monocytes
Cell Type(s) include	Immune cells
Species	Human
Assay / Method	Cell Mitochondrial Stress Test, Glycolysis Stress Test
Cell Seeding Density	4.0x10^6 cells/well
Plate Coating	Not Specified

Authors	Sperry MM, Charrez B, Fotowat H, Gardner E, Pilobello K, Izadifar Z, Lin T, Kuelker A, Kaki S, Lewandowski M, Lightbown S, Martinez R, Marquez S, Moore J, Plaza-Oliver M, Sesay AM, Shcherbina K, Sheehan K, Takeda T, Del Campo D, Andrijauskaite K, Cisneros E, Lopez R, Cano I, Maxwell Z, Jessop I, Veraza R, Bunegin L, Percival TJ, Yracheta J, Pena JJ, Wood DM, Homas ZT, Hinshaw CJ, Cox-Hinshaw J, Parry OG, Sleeter JJ, Weitzel EK, Levin M, Super M, Novak R and Ingber DE
Journal	Elife
Publication Date	24 Sep 2024
Research Area(s) include	Translational Research
Technique	Automated Microscopy and Imaging, Seahorse XF Technology
Analyzer(s) include	BioTek Cytation 5, Seahorse XFe24/XF24
Mode	N/A
Cell Line(s) include	Caco-2, Huh-7, Liver sinusoidal endothelial cells (LSECs), umbilical vein endothelial cells
Cell Type(s) include	Colon cancer cells, Endothelial Cells, Hepatocarcinoma cells
Species	Human
Assay / Method	Cell Mitochondrial Stress Test
Cell Seeding Density	Not Specified
Plate Coating	Not Specified

Authors	N Vannini, V Campos, M Girotra, V Trachsel, S Rojas-Sutterlin, J Tratwal, S Ragusa, E Stefanidis, D Ryu, P Y Rainer, G Nikitin, S Giger, T Y Li, A Semilietof, A Oggier, Y Yersin, L Tauzin, E Pirinen, W C Cheng, J Ratajczak, C Canto, M Ehrbar, F Sizzano, T V Petrova, D Vanhecke, L Zhang, P Romero, A Nahimana, S Cherix, M A Duchosal, P C Ho, B Deplancke, G Coukos, J Auwerx, M P Lutolf, and O Naveiras
Journal	Cell Stem Cell
Publication Date	07 Mar 2019
Research Area(s) include	Food and Nutritional Research, Stem Cell Biology
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XFe24/XF24
Cell Line(s) include	Hematopoietic stem cells
Cell Type(s) include	Stem cells
Species	Mouse
Assay / Method	Cell Mitochondrial Stress Test, Fatty Acid Oxidation
Cell Seeding Density	4.0x10^5 cells/well
Plate Coating	Cell-Tak

Authors	G Eelen, C Dubois, A R Cantelmo, J Goveia, U Bruning, M DeRan, G Jarugumilli, J van Rijssel, G Saladino, F Comitani, A Zecchin, S Rocha, R Chen, H Huang, S Vandekeere, J Kalucka, C Lange, F Morales-Rodriguez, B Cruys, L Treps, L Ramer, S Vinckier, K Brepoels, S Wyns, J Souffreau, L Schoonjans, W H Lamers, Y Wu, J Haustraete, J Hofkens, S Liekens, R Cubbon, B Ghesquiere, M Dewerchin, F L Gervasio, X Li, J D van Buul, X Wu, and P Carmeliet
Journal	Nature
Publication Date	01 Sep 2018
Research Area(s) include	Cardiovascular Research, Cell Physiology Research
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XFe24/XF24
Cell Line(s) include	Endothelial cells
Cell Type(s) include	Endothelial Cells
Species	Human
Assay / Method	Cell Mitochondrial Stress Test
Cell Seeding Density	1.5x10^5 cells/well
Plate Coating	Not Specified

Authors	Feichtinger RG, Olahova M, Kishita Y, Garone C, Kremer LS, Yagi M, Uchiumi T, Jourdain AA, Thompson K, D'Souza AR, Kopajtich R, Alston CL, Koch J, Sperl W, Mastantuono E, Strom TM, Wortmann SB, Meitinger T, Pierre G, Chinnery PF, Chrzanowska-Lightowlers ZM, Lightowlers RN, DiMauro S, Calvo SE, Mootha VK, Moggio M, Sciacco M, Comi GP, Ronchi D, Murayama K, Ohtake A, Rebelo-Guiomar P, Kohda M, Kang D, Mayr JA, Taylor RW, Okazaki Y, Minczuk M, Prokisch H
Journal	Am J Hum Genet
Publication Date	19 Sep 2017
Research Area(s) include	Cardiovascular Research, Cell Physiology Research, Immunology Research
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XFe24/XF24
Cell Line(s) include	MEF
Cell Type(s) include	Neuronal cells
Species	Mouse
Assay / Method	Cell Mitochondrial Stress Test
Cell Seeding Density	2.0x10^4 cells/well
Plate Coating	Not Specified

Authors	L R Fiedler, K Chapman, M Xie, E Maifoshie, M Jenkins, P A Golforoush, M Bellahcene, M Noseda, D Faust, A Jarvis, G Newton, M A Paiva, M Harada, D J Stuckey, W Song, J Habib, P Narasimham, R Aqil, D Sanmugalingam, R Yan, L Pavanello, M Sano, S C Wang, R D Sampson, S Kanayaganam, G E Taffet, L H Michael, M L Entman, T H Tan, S E Harding, C M R Low, C Tralau-Stewart, T Perrior, and M D Schneider
Journal	Cell Stem Cell
Publication Date	04 Apr 2019
Research Area(s) include	Cardiovascular Research, Stem Cell Biology
Technique	Plate Reader-Soluble Sensor, Seahorse XF Technology
Analyzer(s) include	Plate Reader Metabolic Assays-96 well, Seahorse XFe24/XF24
Cell Line(s) include	Cardiac stem cells
Cell Type(s) include	Cardiac cells
Species	Human
Assay / Method	Cell Mitochondrial Stress Test, MitoXpress Intra Intracellular Oxygen Assay
Cell Seeding Density	1.0x10^4 cells/well, 6.0x10^4 cells/well
Plate Coating	Geltrex, Not Specified

Authors	Vandervore LV, Schot R, Milanese C, Smits DJ, Kasteleijn E, Fry AE, Pilz DT, Brock S, Borklu-Yucel E, Post M, Bahi-Buisson N, Sanchez-Soler MJ, van Slegtenhorst M, Keren B, Afenjar A, Coury SA, Tan WH, Oegema R, de Vries LS, Fawcett KA, Nikkels PGJ, Bertoli-Avella A, Al Hashem A, Alwabel AA, Tlili-Graiess K, Efthymiou S, Zafar F, Rana N, Bibi F, Houlden H, Maroofian R, Person RE, Crunk A, Savatt JM, Turner L, Doosti M, Karimiani EG, Saadi NW, Akhondian J, Lequin MH, Kayserili H, van der Spek PJ, Jansen AC, Kros JM, Verdijk RM, Milosevic NJ, Fornerod M, Mastroberardino PG and Mancini GMS
Journal	Am J Hum Genet
Publication Date	05 Dec 2019
Research Area(s) include	Neurobiology Research
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XFe24/XF24
Cell Line(s) include	Dermal Fibroblasts
Cell Type(s) include	Fibroblast
Species	Human
Assay / Method	Cell Mitochondrial Stress Test
Cell Seeding Density	6.0x10^4 cells/well
Plate Coating	Not Specified

Authors	Matsuhashi T, Sato T, Kanno SI, Suzuki T, Matsuo A, Oba Y, Kikusato M, Ogasawara E, Kudo T, Suzuki K, Ohara O, Shimbo H, Nanto F, Yamaguchi H, Saigusa D, Mukaiyama Y, Watabe A, Kikuchi K, Shima H, Mishima E, Akiyama Y, Oikawa Y, Hsin-Jung HO, Akiyama Y, Suzuki C, Uematsu M, Ogata M, Kumagai N, Toyomizu M, Hozawa A, Mano N, Owada Y, Aiba S, Yanagisawa T, Tomioka Y, Kure S, Ito S, Nakada K, Hayashi KI, Osaka H, Abe T
Journal	EBioMedicine
Publication Date	02 June 2017
Research Area(s) include	Neurobiology Research
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XFe24/XF24
Cell Line(s) include	Dermal Fibroblasts
Cell Type(s) include	Fibroblast
Species	Human
Assay / Method	Cell Mitochondrial Stress Test
Cell Seeding Density	Not Specified
Plate Coating	Cell-Tak

Authors	Licht-Mayer S, Campbell GR, Canizares M, Mehta AR, Gane AB, McGill K, Ghosh A, Fullerton A, Menezes N, Dean J, Dunham J, Al-Azki S, Pryce G, Zandee S, Zhao C, Kipp M, Smith KJ, Baker D, Altmann D, Anderton SM, Kap YS, Laman JD, Hart BA, Rodriguez M, Watzlawick R, Schwab JM, Carter R, Morton N, Zagnoni M, Franklin RJM, Mitchell R, Fleetwood-Walker S, Lyons DA, Chandran S, Lassmann H, Trapp BD and Mahad DJ
Journal	Acta Neuropathol
Publication Date	01 Aug 2020
Research Area(s) include	Neurobiology Research
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XFe24/XF24
Cell Line(s) include	Dorsal root ganglion neurons (DRG)
Cell Type(s) include	Neuronal cells
Species	Mouse
Assay / Method	Cell Mitochondrial Stress Test
Cell Seeding Density	Not Specified
Plate Coating	Not Specified

Authors	Y Sun, M Bandi, T Lofton, M Smith, C A Bristow, A Carugo, N Rogers, P Leonard, Q Chang, R Mullinax, J Han, X Shi, S Seth, B A Meyers, M Miller, L Miao, X Ma, N Feng, V Giuliani, M Geck Do, B Czako, W S Palmer, F Mseeh, J M Asara, Y Jiang, P Morlacchi, S Zhao, M Peoples, T N Tieu, M O Warmoes, P L Lorenzi, F L Muller, R A DePinho, G F Draetta, C Toniatti, P Jones, T P Heffernan, and J R Marszalek
Journal	Cell Rep
Publication Date	08 Jan 2019
Research Area(s) include	Cancer Research
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XF 96 well (XF Pro/XFe96/XF96), Seahorse XFe24/XF24, Seahorse XFp
Cell Line(s) include	A549, H1755, H1975, H2122, H460, NB-1, NCI-H1650, PC9, UOK262
Cell Type(s) include	Lung cancer cells, Neuroblastoma, Renal Cell Carcinoma (HLRCC)
Species	Human
Assay / Method	Glycolysis Stress Test
Cell Seeding Density	Not Specified
Plate Coating	Poly-D-lysine and Laminin

Authors	Pua CJ, Tham N, Chin CWL, Walsh R, Khor CC, Toepfer CN, Repetti GG, Garfinkel AC, Ewoldt JF, Cloonan P, Chen CS, Lim SQ, Cai J, Loo LY, Kong SC, Chiang CWK, Whiffin N, de Marvao A, Lio PM, Hii AA, Yang CX, Le TT, Bylstra Y, Lim WK, Teo JX, Padilha K, Silva GV, Pan B, Govind R, Buchan RJ, Barton PJR, Tan P, Foo R, Yip JWL, Wong RCC, Chan WX, Pereira AC, Tang HC, Jamuar SS, Ware JS, Seidman JG, Seidman CE and Cook SA
Journal	Circ Genom Precis Med
Publication Date	20 Aug 2020
Research Area(s) include	Cardiovascular Research
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XF 96 well (XF Pro/XFe96/XF96), Seahorse XFe24/XF24, Seahorse XFp
Cell Line(s) include	iPSC-derived cardiomyocytes
Cell Type(s) include	Cardiac cells
Species	Human
Assay / Method	Basal Metabolic Assay
Cell Seeding Density	Not Specified
Plate Coating	Not Specified

Authors	K Gorgulu, K N Diakopoulos, J Ai, B Schoeps, D Kabacaoglu, A F Karpathaki, K J Ciecielski, E Kaya-Aksoy, D A Ruess, A Berninger, M Kowalska, M Stevanovic, S M Wormann, T Wartmann, Y Zhao, W Halangk, S Voronina, A Tepikin, A M Schlitter, K Steiger, A Artati, J Adamski, M Aichler, A Walch, M Jastroch, G Hartleben, C S Mantzoros, W Weichert, R M Schmid, S Herzig, A Kruger, B Sainz Jr, M Lesina, and H Algul
Journal	Gastroenterology
Publication Date	01 Dec 2018
Research Area(s) include	Cancer Research
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XFe24/XF24
Cell Line(s) include	Tumor cells
Cell Type(s) include	Tumor cells
Species	Mouse
Assay / Method	Cell Mitochondrial Stress Test, Glycolysis Stress Test
Cell Seeding Density	1.0x10^5 cells/well
Plate Coating	Not Specified

Authors	E L Mills, D G Ryan, H A Prag, D Dikovskaya, D Menon, Z Zaslona, M P Jedrychowski, A S H Costa, M Higgins, E Hams, J Szpyt, M C Runtsch, M S King, J F McGouran, R Fischer, B M Kessler, A F McGettrick, M M Hughes, R G Carroll, L M Booty, E V Knatko, P J Meakin, M L J Ashford, L K Modis, G Brunori, D C Sevin, P G Fallon, S T Caldwell, E R S Kunji, E T Chouchani, C Frezza, A T Dinkova-Kostova, R C Hartley, M P Murphy, and L A O'Neill
Journal	Nature
Publication Date	05 Apr 2018
Research Area(s) include	Immunology Research
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XFe24/XF24
Cell Line(s) include	Bone marrow-derived macrophages (BMDM)
Cell Type(s) include	Immune cells
Species	Human
Assay / Method	Glycolysis Stress Test
Cell Seeding Density	2.0x10^5 cells/well
Plate Coating	Not Specified

Authors	Simon J, Nunez-Garcia M, Fernandez-Tussy P, Barbier-Torres L, Fernandez-Ramos D, Gomez-Santos B, Buque X, Lopitz-Otsoa F, Goikoetxea-Usandizaga N, Serrano-Macia M, Rodriguez-Agudo R, Bizkarguenaga M, Zubiete-Franco I, Gutierrez-de Juan V, Cabrera D, Alonso C, Iruzubieta P, Romero-Gomez M, van Liempd S, Castro A, Nogueiras R, Varela-Rey M, Falcon-Perez JM, Villa E, Crespo J, Lu SC, Mato JM, Aspichueta P, Delgado TC and Martinez-Chantar ML
Journal	Cell Metab
Publication Date	03 Mar 2020
Research Area(s) include	Obesity, Diabetes, and Metabolic Disorders
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XFe24/XF24
Cell Line(s) include	Hepatocytes
Cell Type(s) include	Hepatocytes
Species	Mouse
Assay / Method	Fatty Acid Oxidation
Cell Seeding Density	Not Specified
Plate Coating	Not Specified

Authors	G M Fischer, A Jalali, D A Kircher, W C Lee, J L McQuade, L E Haydu, A Y Joon, A Reuben, M P de Macedo, F C L Carapeto, C Yang, A Srivastava, C R Ambati, A Sreekumar, C W Hudgens, B Knighton, W Deng, S D Ferguson, H A Tawbi, I C Glitza, J E Gershenwald, Y N Vashisht Gopal, P Hwu, J T Huse, J A Wargo, P A Futreal, N Putluri, A J Lazar, R J DeBerardinis, J R Marszalek, J Zhang, S L Holmen, M T Tetzlaff, and M A Davies
Journal	Cancer Discov
Publication Date	20 Feb 2019
Research Area(s) include	Cancer Research, Immunology Research, Neurobiology Research
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XF 96 well (XF Pro/XFe96/XF96), Seahorse XFe24/XF24, Seahorse XFp
Cell Line(s) include	Melanoma cells
Cell Type(s) include	Melanoma cells
Species	Human
Assay / Method	Basal Metabolic Assay
Cell Seeding Density	Not Specified
Plate Coating	Not Specified

Authors	J Kalucka, L Bierhansl, N V Conchinha, R Missiaen, I Elia, U Bruning, S Scheinok, L Treps, A R Cantelmo, C Dubois, P de Zeeuw, J Goveia, A Zecchin, F Taverna, F Morales-Rodriguez, A Brajic, L C Conradi, S Schoors, U Harjes, K Vriens, G A Pilz, R Chen, R Cubbon, B Thienpont, B Cruys, B W Wong, B Ghesquiere, M Dewerchin, K De Bock, X Sagaert, S Jessberger, E A V Jones, B Gallez, D Lambrechts, M Mazzone, G Eelen, X Li, S M Fendt, and P Carmeliet
Journal	Cell Metab
Publication Date	04 Dec 2018
Research Area(s) include	Cardiovascular Research
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XFe24/XF24
Cell Line(s) include	Endothelial cells
Cell Type(s) include	Endothelial Cells
Species	Mouse
Assay / Method	Other
Cell Seeding Density	3.0x10^4 cells/well
Plate Coating	Not Specified

Authors	Martinez-Sanchez N, Seoane-Collazo P, Contreras C, Varela L, Villarroya J, Rial-Pensado E, Buque X, Aurrekoetxea I, Delgado TC, Vazquez-Martinez R, Gonzalez-Garcia I, Roa J, Whittle AJ, Gomez-Santos B, Velagapudi V, Tung YCL, Morgan DA, Voshol PJ, Martinez de Morentin PB, Lopez-Gonzalez T, Linares-Pose L, Gonzalez F, Chatterjee K, Sobrino T, Medina-Gomez G, Davis RJ, Casals N, Oresic M, Coll AP, Vidal-Puig A, Mittag J, Tena-Sempere M, Malagon MM, Dieguez C, Martinez-Chantar ML, Aspichueta P, Rahmouni K, Nogueiras R, Sabio G, Villarroya F, Lopez M
Journal	Cell Metab
Publication Date	05 July 2017
Research Area(s) include	Neurobiology Research, Obesity, Diabetes, and Metabolic Disorders, Toxicology Research
Technique	Seahorse XF Technology
Analyzer(s) include	Seahorse XFe24/XF24
Cell Line(s) include	Hepatocytes
Cell Type(s) include	Hepatocytes
Species	Mouse
Assay / Method	Basal Metabolic Assay
Cell Seeding Density	2.0x10^4 cells/well
Plate Coating	Collagen