Gerhard Gstraunthaler

Renal Biochemistry - Molecular Physiology of the Kidney

Epithelial Cell and Tissue Culture

     

Group Members: Gerhard Gstraunthaler, Ph.D., Elisabeth Feifel, Ph.D., Caroline Rauch, Ph.D.,

Research Topics:

Renal Acid-Base Metabolism:

  • Biochemical and Molecular Biological Aspects of Renal Ammoniagenesis and Gluconeogenesis,
  • pH-Sensing and pH-Signaling in Cultured Renal Proximal Tubular Cells,
  • The Role of TGF-b in Renal Acid-Base Adaptation,
  • TGFb-Signaling in Proximal Tubular Cells,

 

Growth, Metabolism, and Differentiation of Epithelial Cells in Tissue Culture:

  • Optimization of Culture Conditions for Human and Animals Cells,
  • Standardization of Cell and Tissue Culture Protocols,
  • Alternatives (Replacement) of Animal Serum in Cell Culture Media,
  • Quality Control and Quality Management (QC/QM) in Cell and Tissue Culture,

Key Words:

  • Acid-Base Homeostasis, Renal Ammoniagenesis, Renal Gluconeogenesis, Ammonia, Glutaminase, PEPCK, pH-Regulated Renal Genes, TGF-b, Cultured Renal Epithelial Cells,

 

General Aspects of Mammalian Cell and Tissue Culture:

  • Serum-free Cell and Tissue Culture,
  • The Serum-free Media Interactive Online Database – see below,
  • Alternatives (Replacement) of Animal Serum in Cell Culture Media,
  • Quality Control and Quality Management (QC/QM) in Cell and Tissue Culture,

 

Textbook on Cell and Tissue Culture (“Zell- und Gewebekultur”) – see below,

International Collaboration:

  • Norman P. Curthoys, Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO 80523-1870, USA,

 

  • ECVAM, European Centre for the Validation of Alternative Methods, JRC, Joint Research Centre of the European Commission, Ispra, Italy (ecvam.jrc.it)

 

 

 

Selected Publications:

Gstraunthaler G. et al.. Biochemical characterization of renal epithelial cell cultures (LLC-PK1 and MDCK). Am. J. Physiol. 248: F536-F544 (1985).

Gstraunthaler G. and Handler J.S. Isolation, growth, and characterization of a gluconeogenic strain of renal cells. Am. J. Physiol. 252: C232-C238 (1987).

Gstraunthaler G.J.A. Epithelial cells in tissue culture (Review). Renal Physiol. Biochem. 11: 1-42 (1988).

Gstraunthaler G. et al. Morphological and biochemical changes of LLC-PK1 cells during adaptation to glucose-free culture conditions. Renal Physiol. Biochem. 13: 137-153 (1990).

Gstraunthaler G. et al. Renal cell cultures: A tool for studying tubular function and nephrotoxicity. Toxicol. Lett. 53: 1-7 (1990).

Gstraunthaler G. et al. Ammoniagenesis in LLC-PK1 cultures: role of transamination. Am. J. Physiol. 263: C47-C54 (1992).

Gstraunthaler G. et al. Ammoniagenesis in renal cell culture. Lack of extracellular ammoniagenesis at the apical surface of LLC-PK1 epithelia. Renal Physiol. Biochem. 16: 203-211 (1993).

Gstraunthaler G. et al. A novel gluconeogenic strain of OK cells with metabolic properties different from gluconeogenic LLC-PK1 cells. Cell. Physiol. Biochem. 3: 78-88 (1993).

Netzer A. and Gstraunthaler G. Selective release of apical membrane enzymes from cultured renal epithelia by phosphatidylinositol-specific phospholipase C. Renal Physiol. Biochem. 16: 299-310 (1993).

Gstraunthaler G.J.A. Ammoniagenesis in renal cell culture. A comparative study on ammonia metabolism of renal epithelial cell lines. Contrib. Nephrol. 110: 88-97 (1994).

Holcomb T. et al. Subcellular localization of PEPCK and metabolism of gluconeogenic substrains of renal cell lines. Am. J. Physiol. 268: C449-C457 (1995).

Liu W. et al. PMA and staurosporine affect expression of the PCK gene in LLC-PK1-F+ cells. Am. J. Physiol. 275: F361-F369 (1998).

Gstraunthaler G. et al. Impact of culture conditions, culture media volumes, and glucose content on metabolic properties of renal epithelial cell cultures. Are renal cells in tissue culture hypoxic? Cell. Physiol. Biochem. 9: 150-172 (1999).

Gstraunthaler G. et al. Differential expression and acid-base regulation of glutaminase mRNAs in gluconeogenic LLC-PK1-FBPase+ cells. Am. J. Physiol. Renal Physiol. 278: F227-F237 (2000).

Curthoys N.P. and Gstraunthaler G. Mechanism of increased renal gene expression during metabolic acidosis (Invited Review). Am. J. Physiol. Renal Physiol. 281: F381-F390 (2001).

Hartung T. et al. Good Cell Culture Practice. ECVAM Good cell culture practice task force report 1. ATLA 30: 407-414 (2002).

Feifel E. et al. p38 MAPK mediates acid-induced transcription of PEPCK in LLC-PK1-FBPase+ cells. Am. J. Physiol. Renal Physiol. 283: F678-F688 (2002).

Gstraunthaler G. Alternatives to the use of fetal bovine serum: Serum-free cell culture (Review). ALTEX 20: 275-281 (2003).

van der Valk J. et al. The humane collection of fetal bovine serum and possibilities for serum-free cell and tissue culture. Toxicol. in Vitro 18: 1-12 (2004).

Coecke S. et al. Guidance on Good Cell Culture Practice. A report of the second ECVAM task force on Good Cell Culture Practice. ATLA 33: 261-287 (2005).

O´Hayre M. et al. Effects of constitutively active and dominant negative MAPK kinase (MKK) 3 and MKK6 on the pH-repsonsive increase in phosphoenolpyruvate carboxykinase mRNA. J. Biol. Chem. 281: 2982-2988 (2006).

Balls M. et al. The Importance of Good Cell Culture Practice (GCCP). ALTEX 23 (Special Issue): 270-273 (2006).

Gstraunthaler G. Standardization in Cell and Tissue Culture - The Need for Specific GLP Guidelines in the Cell Culture Laboratory (Good Cell Culture Practice - GCCP). ALTEX 23 (Special Issue): 274-277 (2006).

Andratsch M .et al.  TGF-β signaling and its effect on glutaminase expression in LLC-PK1-FBPase+ cells. Am. J. Physiol. Renal Physiol. 293: F846-F853 (2007).

Brunner D. et al. Serum-free Cell Culture: The Serum-free Media Interactive Online Database. ALTEX 27: 53-62 (2010).

van der Valk J. et al. Optimization of chemically defined cell culture media – Replacing fetal bovine serum in mammalian in vitro methods. Toxicol. in Vitro 24: 1053-1063 (2010).

Gstraunthaler G. The Bologna Statement on Good Cell Culture Practice (GCCP) – 10 years later. ALTEX 27 (Special Issue): 141-146 (2010).

Rauch C. et al. Alternatives to the Use of Fetal Bovine Serum: Human Platelet Lysates as a Serum Substitute in Cell Culture Media. ALTEX 28: 305-316 (2011).

 


Recent Abstracts:

Gstraunthaler G. Safety in the Cell Culture Laboratory. 13. Kongreß über Alternativen zu Tierversuchen, Linz, Austria. ALTEX 23: 95, 2006.

Tauß T. et al. Dramatic increase in numbers of transgenic mice – we must take action now. 14th Congress on Alternatives to Animal Testing, Linz, Austria. ALTEX 24: 230-231, 2007.

Gstraunthaler G. Biological differences between embryonic and adult stem cells. 15th Congress on Alternatives to Animal Testing, Linz, Austria, ALTEX 25 (Suppl. 1): 23, 2008.

Gstraunthaler G. et al. Alternatives to the use of fetal bovine serum (FBS): Recent strategies to reduce or replace FBS in cell and tissue culture. 15th Congress on Alternatives to Animal Testing, Linz, Austria. ALTEX 25 (Suppl. 1): 24, 2008.

Rauch C. et al. Human Platelet Lysates as a Serum Substitute in Renal Epithelial Cell Culture. Annual Meeting of the American Society of Nephrology, Denver, CO, USA, 2010.

Rauch C. et al. Use of human platelet lysates in stem cell-based alternative testing strategies. ÖGMBT Annual Meeting 2011, Salzburg, Austria.

  www.stiftung-set.de

 

 

         New Edition / Neuauflage:

            Textbook on   Cell and Tissue Culture   (in German):

                                                           Toni Lindl und Gerhard Gstraunthaler

                                                           Zell- und Gewebekultur

                                                           Von den Grundlagen zur Laborbank

                                                           6. Aufl., Spektrum Akademischer Verlag, Heidelberg

                                                           ISBN: 978-3-8274-1776-3

            Link:  http://www.springer.com/spektrum+akademischer+verlag/spektrum-sachb%C3%BCcher/biologie/book/978-3-8274-1776-3

 

 

FETAL BOVINE SERUM IN CELL AND TISSUE CULTURE

 

The interactive online database for serum-free cell culture media

 

 

            Fetal bovine serum (FBS) is commonly used as an essential supplement to cell culture media. FBS is a cocktail of most of the factors required for cell attachment, growth, and proliferation in vitro.

 

            DISADVANTAGES IN THE USE OF FBS

 

            However, the use of animal serum also bears a number of disadvantages.

These can either be seen from

            (a) a theoretical, cell biological point of view, since serum in general is ill-defined,

            (b) from ethical perspectives in terms of animal protection arguments about harvest and collection of FBS from bovine fetuses, and

            (c) in terms of recent concerns about the global supply vs. demand of FBS.

 

            It is estimated that about 500.000 litres FBS are produced per year for the world market. This means, that more than 1.000.000 bovine fetuses have to be harvested, and it is expected, that these numbers will continue to increase annually. As a consequence, in terms of the 3Rs, a number of strategies have been developed to reduce or replace the requirement for FBS in cell culture media. As a major goal of these initiatives, any efforts shall be undertaken in order to decrease the global demands for FBS and thus to decrease the number of bovine fetuses needed.

 

            ALTERNATIVES TO FETAL BOVINE SERUM

 

            During the last three decades, FBS could be substituted through other supplements or by the use of defined chemical components in serum-free cell culture. A number of serum-free media formulations have been described for continuous mammalian and insect cell lines as well as for primary cultures. However, switching to serum-free media still needs a time-consuming literature survey and manufacturer search for appropriate media formulations, respectively.

 

            THE SERUM-FREE DATABANK

 

            In a free accessible, interactive online database (www.goodcellculture.com) commercially available serum-free media and continuous cell lines already adapted to serum-free culture can be searched for by means of different criteria. The serum-free media interactive online database is described in detail in ALTEX 27(1): 53-62, 2010.

 

            Searchable criteria are

            •  the degree of chemical definition, e.g. serum-free, animal-derived component-free or chemically defined, and

            •  the kind of medium, e.g. basal media, media supplements, or full replacement media.

 

            In order to specify the cell lines that are adapted for serum-free media, search terms like organism, organ, tissue, cell type and disease can be used. All commercially available serum-free media and adapted cell lines currently available from main distributors (e.g. ATCC, ECACC and DMSZ) are also included in the database.

 

 

SEARCH THE INTERACTIVE ONLINE DATABASE FOR SERUM-FREE CELL CULTURE MEDIA AT:

www.goodcellculture.com

 

 

updated: December 9, 2011

 

E-Mail to gerhard.gstraunthaler@i-med.ac.at

Division of Physiology, Innsbruck Medical University

Fritz-Pregl-Strasse 3, A-6020 INNSBRUCK, AUSTRIA

Phone: ++43 (512) 9003-70810, FAX: ++43 (512) 9003-73800

Web Site:  http://physiologie.i-med.ac.at/

 

Impressum:  Department of Physiology and Medical Physics

 

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