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Article: Preparation of a universally usable, animal product free, defined medium for 2D and 3D culturing of normal and cancer cells.

Weber, Tilo / Bajramovic, Jeffrey / Oredsson, Stina

2024 Volume 12, Page(s) 102592

Abstract: Since 1958, cell culture media supplemented with fetal bovine serum is used, despite the well-known concerns about animal welfare, reproducibility, reliability, relevance, and safety. To obliterate these concerns and increase scientific accuracy, we ... ...

Abstract	Since 1958, cell culture media supplemented with fetal bovine serum is used, despite the well-known concerns about animal welfare, reproducibility, reliability, relevance, and safety. To obliterate these concerns and increase scientific accuracy, we recently published an open access, publicly available paper on a defined medium composition to make it possible for any lab to prepare this medium. The medium supports routine culturing and cell banking as well as investigations of growth curves, dose response testing of compounds of cells in 2D and 3D, and cell migration; all important aspects for research and toxicology. Here we give a detailed description of how to mix the defined universal cell culture medium in 14 simple steps to support any entity that wishes to make it. We also list different normal and cancer cell lines that have been cultured in the defined medium.•Open source composition of animal product free universal cell culture medium•Protocols for mixing solutions of small xeno free molecules for supplementation•Protocols for mixing solutions of human proteins for supplementation.
Language	English
Publishing date	2024-02-03
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	2830212-6
ISSN	2215-0161
ISSN	2215-0161
DOI	10.1016/j.mex.2024.102592
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Article ; Online: Have the non-technical summaries of animal experiments in Europe improved? An update.

Taylor, Katy / Weber, Tilo / Alvarez, Laura Rego

ALTEX

2024

Abstract: Following a review of the Directive 2010/63/EU on the protection of animals used for scientific purposes in the European Union (EU), non-technical project summaries (NTS) of all approved projects must be published in a central database using a standard ... ...

Abstract	Following a review of the Directive 2010/63/EU on the protection of animals used for scientific purposes in the European Union (EU), non-technical project summaries (NTS) of all approved projects must be published in a central database using a standard template. Our initial review of the NTS reported in ALTEX in 2018 had found the NTS to be deficient in their accessibility and quality, notably the adverse effects section where the harms to the animals are meant to be described. Here we repeat our review to see if these legislative changes have improved the accessibility and quality of the NTS. As before we focused on the NTS from the United Kingdom (UK) and Germany; even though the UK has left the EU it is using the same template. We found significant improvement in the reporting of five of the six elements we identified as essential to the predicted harms section. However, there was no significant improvement in the reporting of adverse effects. Only 41% of German NTS and 48% of UK NTS are fully reporting this important element of the predicted harms section. In our view, researchers need support in describing the impact of their research on the animals and to assist here we include a checklist for competent authorities and a list of suggested terminology for standard administration and sampling procedures. Unless the NTS improve further, their utility as a tool for sharing of good practices in the 3Rs or to support evidence-based policy making will remain limited.
Language	English
Publishing date	2024-03-14
Publishing country	Germany
Document type	Journal Article
ZDB-ID	165707-0
ISSN	1868-8551 ; 1018-4562 ; 0946-7785
ISSN (online)	1868-8551
ISSN	1018-4562 ; 0946-7785
DOI	10.14573/altex.2310181
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Article: The ethical evaluation of animal experiments deserves more than empty phrases.

Weber, Tilo

ALTEX

2018 Volume 35, Issue 4, Page(s) 518–519

MeSH term(s)	Animal Experimentation/ethics ; Animal Experimentation/legislation & jurisprudence ; Animal Experimentation/standards ; Animal Welfare ; Animals ; Ethics, Research ; Germany
Language	English
Publishing date	2018-12-18
Publishing country	Germany
Document type	Letter
ZDB-ID	165707-0
ISSN	1868-596X ; 1018-4562 ; 0946-7785
ISSN	1868-596X ; 1018-4562 ; 0946-7785
DOI	10.14573/altex.1810041
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Article: Replacing fetal bovine serum (FBS) in research and testing.

Weber, Tilo / Wagner, Kristina

ALTEX

2021 Volume 38, Issue 1, Page(s) 163–164

MeSH term(s)	Cells, Cultured ; Culture Media ; Fetal Blood ; Serum ; Serum Albumin, Bovine
Chemical Substances	Culture Media ; Serum Albumin, Bovine (27432CM55Q)
Language	English
Publishing date	2021-01-16
Publishing country	Germany
Document type	Editorial
ZDB-ID	165707-0
ISSN	1868-596X ; 1018-4562 ; 0946-7785
ISSN	1868-596X ; 1018-4562 ; 0946-7785
DOI	10.14573/altex.2012141
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Article: Case Studies Exemplifying the Transition to Animal Component-free Cell Culture.

Weber, Tilo / Wiest, Joachim / Oredsson, Stina / Bieback, Karen

Alternatives to laboratory animals : ATLA

2022 Volume 50, Issue 5, Page(s) 330–338

Abstract: Cell culture techniques are strongly connected with modern scientific laboratories and production facilities. Thus, choosing the most suitable medium for the cells involved is vital, not only directly to optimise cell viability but also indirectly to ... ...

Abstract	Cell culture techniques are strongly connected with modern scientific laboratories and production facilities. Thus, choosing the most suitable medium for the cells involved is vital, not only directly to optimise cell viability but also indirectly to maximise the reliability of the experiments performed with the cells. Fetal bovine or calf serum (FBS or FCS, respectively) is the most commonly used cell culture medium supplement, providing various nutritional factors and macromolecules essential for cell growth. Yet, the use of FBS encompasses a number of disadvantages. Scientifically, one of the most severe disadvantages is the lot-to-lot variability of animal sera that hampers reproducibility. Therefore, transitioning from the use of these ill-defined, component-variable, inconsistent, xenogenic, ethically questionable and even potentially infectious media supplements, is key to achieving better data reproducibility and thus better science. To demonstrate that the transition to animal component-free cell culture is possible and achievable, we highlight three different scenarios and provide some case studies of each, namely: i) the adaptation of single cell lines to animal component-free culture conditions by the replacement of FBS and trypsin; ii) the adaptation of multicellular models to FBS-free conditions; and (iii) the replacement of FBS with human platelet lysate (hPL) for the generation of primary stem/stromal cell cultures for clinical purposes. By highlighting these examples, we aim to foster and support the global movement towards more consistent science and provide evidence that it is indeed possible to step out of the currently smouldering scientific reproducibility crisis.
MeSH term(s)	Animals ; Cattle ; Cell Culture Techniques/methods ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Humans ; Mesenchymal Stem Cells ; Reproducibility of Results ; Trypsin
Chemical Substances	Trypsin (EC 3.4.21.4)
Language	English
Publishing date	2022-08-19
Publishing country	England
Document type	Journal Article
ZDB-ID	605800-0
ISSN	0261-1929
ISSN	0261-1929
DOI	10.1177/02611929221117999
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Article ; Online: Practical workshop on replacing fetal bovine serum (FBS) in life science research: From theory into practice.

Eggert, Sebastian / Wiest, Joachim / Rosolowski, Jessica / Weber, Tilo

ALTEX

2022 Volume 39, Issue 4, Page(s) 712–713

MeSH term(s)	Animals ; Serum Albumin, Bovine ; Animal Testing Alternatives ; Culture Media, Serum-Free ; Culture Media ; Biological Science Disciplines ; Serum ; Cells, Cultured
Chemical Substances	Serum Albumin, Bovine (27432CM55Q) ; Culture Media, Serum-Free ; Culture Media
Language	English
Publishing date	2022-10-18
Publishing country	Germany
Document type	Editorial
ZDB-ID	165707-0
ISSN	1868-8551 ; 1018-4562 ; 0946-7785
ISSN (online)	1868-8551
ISSN	1018-4562 ; 0946-7785
DOI	10.14573/altex.2207071
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Article: Case Studies Exemplifying the Transition to Animal Component-free Cell Culture

Weber, Tilo / Wiest, Joachim / Oredsson, Stina / Bieback, Karen

ATLA. Alternatives to laboratory animals. ,

2022

Abstract	Cell culture techniques are strongly connected with modern scientific laboratories and production facilities. Thus, choosing the most suitable medium for the cells involved is vital, not only directly to optimise cell viability but also indirectly to maximise the reliability of the experiments performed with the cells. Fetal bovine or calf serum (FBS or FCS, respectively) is the most commonly used cell culture medium supplement, providing various nutritional factors and macromolecules essential for cell growth. Yet, the use of FBS encompasses a number of disadvantages. Scientifically, one of the most severe disadvantages is the lot-to-lot variability of animal sera that hampers reproducibility. Therefore, transitioning from the use of these ill-defined, component-variable, inconsistent, xenogenic, ethically questionable and even potentially infectious media supplements, is key to achieving better data reproducibility and thus better science. To demonstrate that the transition to animal component-free cell culture is possible and achievable, we highlight three different scenarios and provide some case studies of each, namely: i) the adaptation of single cell lines to animal component-free culture conditions by the replacement of FBS and trypsin; ii) the adaptation of multicellular models to FBS-free conditions; and (iii) the replacement of FBS with human platelet lysate (hPL) for the generation of primary stem/stromal cell cultures for clinical purposes. By highlighting these examples, we aim to foster and support the global movement towards more consistent science and provide evidence that it is indeed possible to step out of the currently smouldering scientific reproducibility crisis.
Keywords	blood serum ; calves ; cell culture ; cell growth ; cell viability ; culture media ; humans ; laboratories ; trypsin
Language	English
Dates of publication	2022-0819
Publishing place	SAGE Publications
Document type	Article
ZDB-ID	605800-0
ISSN	0261-1929
ISSN	0261-1929
DOI	10.1177/02611929221117999
Database	NAL-Catalogue (AGRICOLA)

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Article: Reply to comment "Animal welfare and ethics in the collection of fetal blood for the production of fetal bovine serum".

Weber, Tilo / Wirths, Frigga / Brakebusch, Nina / Van der Valk, Jan

ALTEX

2021 Volume 38, Issue 2, Page(s) 324–326

MeSH term(s)	Animal Welfare ; Animals ; Fetal Blood ; Serum Albumin, Bovine
Chemical Substances	Serum Albumin, Bovine (27432CM55Q)
Language	English
Publishing date	2021-04-19
Publishing country	Germany
Document type	Editorial ; Comment
ZDB-ID	165707-0
ISSN	1868-596X ; 1018-4562 ; 0946-7785
ISSN	1868-596X ; 1018-4562 ; 0946-7785
DOI	10.14573/altex.2103191
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Article ; Online: A new animal product free defined medium for 2D and 3D culturing of normal and cancer cells to study cell proliferation and migration as well as dose response to chemical treatment

Rafnsdóttir, Ólöf Birna / Kiuru, Anna / Tebäck, Mattis / Friberg, Nathalie / Revstedt, Philippa / Zhu, Johan / Thomasson, Sofia / Czopek, Agnieszka / Malakpour-Permlid, Atena / Weber, Tilo / Oredsson, Stina

Toxicology Reports. 2023, v. 10 p.509-520

2023

Abstract: Cell culturing methods are increasingly used to reduce and replace the use of live animals in biomedical research and chemical toxicity testing. Although live animals are avoided when using cell culturing methods, they often contain animal-derived ... ...

Abstract	Cell culturing methods are increasingly used to reduce and replace the use of live animals in biomedical research and chemical toxicity testing. Although live animals are avoided when using cell culturing methods, they often contain animal-derived components of which one of the most commonly used is foetal bovine serum (FBS). FBS is added to cell culture media among other supplements to support cell attachment/spreading and cell proliferation. The safety, batch-to-batch variation, and ethical problems with FBS are acknowledged and therefore world-wide efforts are ongoing to produce FBS free media. Here, we present the composition of a new defined medium with only human proteins either recombinant or derived from human tissues. This defined medium supports long-term culturing/routine culturing of normal cells and of cancer cells, and can be used for freezing and thawing of cells, i.e. for cell banking. Here, we show for our defined medium, growth curves and dose response curves of cells grown in two and three dimensions, and applications such as cell migration. Cell morphology was studied in real time by phase contrast and phase holographic microscopy time-lapse imaging. The cell lines used are human cancer-associated fibroblasts, keratinocytes, breast cancer JIMT-1 and MDA-MB-231 cells, colon cancer CaCo-2 cells, and pancreatic cancer MiaPaCa-2 cells as well as the mouse L929 cell line. In conclusion, we present the composition of a defined medium without animal-derived products which can be used for routine culturing and in experimental settings for normal cells and for cancer cells, i.e. our defined medium provides a leap towards a universal animal product free cell culture medium.
Keywords	animal products ; biomedical research ; breast neoplasms ; cell culture ; cell lines ; cell movement ; cell proliferation ; cell structures ; chemical treatment ; colorectal neoplasms ; culture media ; dose response ; ethics ; fetal bovine serum ; fibroblasts ; humans ; keratinocytes ; mice ; microscopy ; pancreatic neoplasms ; toxicity ; toxicology ; 2D ; 3D ; CAFs ; DAPI ; hi-DHHS ; FBS ; hi-FBS ; GIVIMP ; PCL ; PBS ; SAEC ; Animal product free defined medium ; Cancer cells ; Normal cells ; Routine culturing ; Growth curves ; Dose response curves ; Cell migration ; Time-lapse imaging ; Two dimensions ; Three dimensions
Language	English
Size	p. 509-520.
Publishing place	Elsevier B.V.
Document type	Article ; Online
Note	Pre-press version ; Use and reproduction
ZDB-ID	2805786-7
ISSN	2214-7500
ISSN	2214-7500
DOI	10.1016/j.toxrep.2023.04.001
Database	NAL-Catalogue (AGRICOLA)

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Article: A worldwide survey on the use of animal-derived materials and reagents in scientific experimentation.

Cassotta, Manuela / Bartnicka, Joanna Julia / Pistollato, Francesca / Parvatam, Surat / Weber, Tilo / D'Alessandro, Vito / Bastos, Luisa Ferreira / Coecke, Sandra

Engineering in life sciences

2022 Volume 22, Issue 9, Page(s) 564–583

Abstract: The use of cell and tissue-based methods in basic, applied and regulatory science has been increasing exponentially. Animal-derived components, including serum, coating materials, growth factors and antibodies are routinely used in cell/tissue cultures ... ...

Abstract	The use of cell and tissue-based methods in basic, applied and regulatory science has been increasing exponentially. Animal-derived components, including serum, coating materials, growth factors and antibodies are routinely used in cell/tissue cultures and in general laboratory practices. In addition to ethical issues, the use and production of animal-derived materials and reagents raises many scientific concerns, generally associated with presence of undefined components and batch-to-batch variability, which may compromise experimental reproducibility. On the other hand, non-animal materials and reagents, such as human cells, alternatives to animal sera or non-animal recombinant antibodies, are becoming increasingly available, and their use is encouraged by the EU Directive 2010/63 and the Guidance Document on Good In vitro Method Practices (GIVIMP), published by the Organization for Economic Cooperation and Development (OECD). In an effort to map the current state of use of animal-derived reagents across different sectors and to identify the obstacles possibly hampering the implementation of non-animal derived alternatives, a global online survey addressed to scientists working on in vivo, in vitro, in silico methods, in academia as well as pharmaceutical or cosmetic companies, was conducted with the goal to understand: 1) the most commonly used animal-derived materials and reagents, 2) the main issues associated with the production and use of animal-derived materials and reagents, 3) the current level of knowledge on available non-animal alternative materials and reagents, and 4) what educational and information sources could be most useful or impactful to disseminate knowledge on non-animal alternatives. This paper provides an overview of the survey replies and discusses possible proposals to increase awareness, acceptance and use of non-animal ingredients.
Language	English
Publishing date	2022-07-18
Publishing country	Germany
Document type	Journal Article
ZDB-ID	2071199-2
ISSN	1618-2863 ; 1618-0240
ISSN (online)	1618-2863
ISSN	1618-0240
DOI	10.1002/elsc.202100167
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