Standardization in Cell and Tissue Culture - The Need for Specific GLP Guidelines in the Cell Culture Laboratory (Good Cell Culture Practice - GCCP)

G Gstraunthaler¹, S Coecke², M Balls³, G Bowe², J Davis⁴, T Hartung², R Hay⁵, O-W Merten⁶, A Price², WS Stokes⁷, LM Schechtman⁸, G Stacey⁹

1. Dept. of Physiology and Medical Physics, Innsbruck Medical University, Innsbruck, Austria
2. ECVAM, Institute for Health & Consumer Protection, European Commission Joint Research Centre, Ispra, Italy
3. FRAME, Russell & Burch House, Nottingham, UK
4. Research and Development Dept., Bio-Products Laboratory, Herts., UK
5. ATCC, Manassas, Virginia, USA
6. Généthon, Evry, France
7. NIEHS, NICEATM, RTP, NC, USA
8. National Center for Toxicological Research, U.S. FDA, Rockville, MD, USA
9. National Institute for Biological Standards and Control, Division of Cell Biology and UK Stem Cell Bank, Herts., UK

The cultivation of eukaryotic cells has become a powerful technique in basic cell and molecular biological research, applied biotechnology, and in vitro alternatives. Before cell culture could be carried out successfully, two problems had to be overcome:

1. Populations of cells had to be established from single cells; and
2. these populations had to be maintained for many generations.
   
   

In a successful propagation of cells in vitro, cells from various tissues should grow and proliferate under appropriate culture conditions, while preserving highly differentiated functions, which closely resemble their ancestor cells in vivo. Thus, cell proliferation and cell differentiation are two major, albeit opposing, end points in tissue culture. Which of these contrasting goals should be achieved depends on the aim of a selected cell culture study and thus, on the culture conditions applied:

1. the supplementation of culture media with growth factors or differentiation factors,
2. the use of specific extracellular matrix components,
3. the subcultivation intervals and seeding densities,
4. the feeding cycles, and
5. stationary cultures versus dynamic media supply in perfusion reactors.
   
   

In sum, a number of tissue culture parameters have to be defined and coordinated. However, despite the widespread use and broad applications of cell and tissue cultures, a significant number of basic questions and methodological protocols are still unsolved and are handled in various ways by tissue culture laboratories. Selected examples will be presented, on how culture medium composition, medium volumes, feeding cycles, serum supplementation, or use of extracellular matrix components will influence growth of cultured cells and the expression of differentiated functions, which represents a serious impact on the credibility, reliability, reproducibility, and comparablity of in vitro alternatives.

In conclusion, a minimum set of standards has to be defined in order to establish reproducibility and interlaboratory comparability of results obtained with in vitro cell culture technologies. Therefore, in analogy to GLP, a Good Cell Culture Practice (GCCP), i. e. good laboratory practice in the cell culture laboratory, was initiated at the 3rd World Congress on Alternatives and Animal Use in the Life Sciences in Bologna, 1999. As a result, GCCP Guidelines were elaborated by an ECVAM Task Force and published in ATLA (vol. 30: 407-414, 2002). Following this publication, a new GCCP Task Force was convened at ECVAM, Ispra, Italy, in order to produce an updated GCCP Guidance document (ATLA 33: 261-287, 2005), which will be presented in an accompanying lecture.

Date: Tuesday, August 23, 2005, 14.00Ð16.00 h, Estrel Hall C5/C6

5.7 Session: Progress in Quality Assurance for In Vitro Alternative Studies