R. Ian Freshney

Bio: R. Ian Freshney is an academic researcher from University of Glasgow. The author has contributed to research in topics: Cell culture & Cellular differentiation. The author has an hindex of 16, co-authored 40 publications receiving 4132 citations.

Animal cell culture : a practical approach

Abstract: R.I. Freshney: Introduction to basic principles H.R. Maurer: Towards serum-free, chemically defined media for mammalian cell culture B. Griffiths: Scaling-up of animal cell cultures R.J. Hay: Cell line preservation and characterization D. Conkie: Separation of viable cells by centrifugal elutriation J.V. Watson & E. Erba: Flow cytometry I. Lasnitzki: Organ culture A.P. Wilson: Cytotoxicity and viability assays S.M. Lang, A.H. Wyllie, & D. Conkie: In situ hybridization Index.

Check your cultures! A list of cross-contaminated or misidentified cell lines.

Abstract: Continuous cell lines consist of cultured cells derived from a specific donor and tissue of origin that have acquired the ability to proliferate indefinitely. These cell lines are well-recognized models for the study of health and disease, particularly for cancer. However, there are cautions to be aware of when using continuous cell lines, including the possibility of contamination, in which a foreign cell line or microorganism is introduced without the handler's knowledge. Cross-contamination, in which the contaminant is another cell line, was first recognized in the 1950s but, disturbingly, remains a serious issue today. Many cell lines become cross-contaminated early, so that subsequent experimental work has been performed only on the contaminant, masquerading under a different name. What can be done in response—how can a researcher know if their own cell lines are cross-contaminated? Two practical responses are suggested here. First, it is important to check the literature, looking for previous work on cross-contamination. Some reports may be difficult to find and to make these more accessible, we have compiled a list of known cross-contaminated cell lines. The list currently contains 360 cell lines, drawn from 68 references. Most contaminants arise within the same species, with HeLa still the most frequently encountered (29%, 106/360) among human cell lines, but interspecies contaminants account for a small but substantial minority of cases (9%, 33/360). Second, even if there are no previous publications on cross-contamination for that cell line, it is essential to check the sample itself by performing authentication testing.

Culture of epithelial cells

Abstract: Contributors. Preface (R. Freshney & M. Freshney). Preface to the First Edition (R. Freshney). List of Abbreviations. Introduction (R. Freshney). Cell Interaction and Epithelial Differentiation (N. Maas Szabowski, et al.). The Epidermis (E. Parkinson & W. Yeudall). Culture of Human Mammary Epithelial Cells (M. Stampfer, et al.). Culture of Human Cervical Epithelial Cells (M. Stanley). Human Prostatic Epithelial Cells (D. Peehl). Human Oral Epithelium (R. Grafstrom). Normal Human Bronchial Epithelial Cell Culture (J. Wise & J. Lechner). Isolation and Culture of Pulmonary Alveolar Epithelial Type II Cells (L. Dobbs & R. Gonzalez). Isolation and Culture of Intestinal Epithelial Cells (C. Booth & J. O Shea). Isolation and Culture of Animal and Human Hepatocytes (C. Guguen Guillouzo). Culture of Human Urothelium (J. Southgate, et al.). Other Epithelial Cells (R. Freshney). List of Suppliers. Index.

Culture of Cells for Tissue Engineering

Abstract: Preface. List of Abbreviations. PART I: CELL CULTURE. 1. Basic Principles of Cell Culture (R. Freshney). 2. Mesenchymal Stem Cells for Tissue Engineering (D. Lennon & A. Caplan). 3. Human Embryonic Stem cell Culture for Tissue Engineering (S. Levenberg, et al.). 4. Cell Sources for Cartilage Tissue Engineering (B. Johnstone, et al.). 5. Lipid-Mediated Gene Transfer for Cartilage Tissue Engineering (H. Madry). PART II: TISSUE ENGINEERING. 6. Tissue Engineering: Basic Considerations (G. Vunjak-Novakovic). 7. Tissue Engineering of Articular Cartilage (K. Masuda & R. Sah). 8. Ligament Tissue Engineering (J. Chen, et al.). 9. Cellular Photoencapsulation in Hydrogels (J. Elisseeff, et al.). 10. Tissue Engineering Human Skeletal Muscle for Clinical Applications (J. Shansky, et al.). 11. Engineered Heart Tissue (T. Eschenhagen & W. Zimmermann). 12. Tissue-Engineered Blood Vessels (R. Klinger & L. Niklason). 13. Tissue Engineering of Bone (S. Hofmann, et al.). 14. Culture of Neuroendocrine and Neuronal Cells for Tissue Engineering (P. Lelkes, et al.). 15. Tissue Engineering of the Liver (G. Underhill, et al.). Suppliers List. Glossary. Index.

Adult pancreatic beta-cells are formed by self-duplication rather than stem-cell differentiation.

Abstract: How tissues generate and maintain the correct number of cells is a fundamental problem in biology. In principle, tissue turnover can occur by the differentiation of stem cells, as is well documented for blood, skin and intestine, or by the duplication of existing differentiated cells. Recent work on adult stem cells has highlighted their potential contribution to organ maintenance and repair. However, the extent to which stem cells actually participate in these processes in vivo is not clear. Here we introduce a method for genetic lineage tracing to determine the contribution of stem cells to a tissue of interest. We focus on pancreatic beta-cells, whose postnatal origins remain controversial. Our analysis shows that pre-existing beta-cells, rather than pluripotent stem cells, are the major source of new beta-cells during adult life and after pancreatectomy in mice. These results suggest that terminally differentiated beta-cells retain a significant proliferative capacity in vivo and cast doubt on the idea that adult stem cells have a significant role in beta-cell replenishment.

Spheroid-based drug screen: considerations and practical approach

Abstract: Although used in academic research for several decades, 3D culture models have long been regarded expensive, cumbersome and unnecessary in drug development processes. Technical advances, coupled with recent observations showing that gene expression in 3D is much closer to clinical expression profiles than those seen in 2D, have renewed attention and generated hope in the feasibility of maturing organotypic 3D systems to therapy test platforms with greater power to predict clinical efficacies. Here we describe a standardized setup for reproducible, easy-handling culture, treatment and routine analysis of multicellular spheroids, the classical 3D culture system resembling many aspects of the pathophysiological situation in human tumor tissue. We discuss essential conceptual and practical considerations for an adequate establishment and use of spheroid-based drug screening platforms and also provide a list of human carcinoma cell lines, partly on the basis of the NCI-DTP 60-cell line screen, that produce treatable spheroids under identical culture conditions. In contrast to many other settings with which to achieve similar results, the protocol is particularly useful to be integrated into standardized large-scale drug test routines as it requires a minimum number of defined spheroids and a limited amount of drug. The estimated time to run the complete screening protocol described herein--including spheroid initiation, drug treatment and determination of the analytical end points (spheroid integrity, and cell survival through the acid phosphatase assay)--is about 170 h. Monitoring of spheroid growth kinetics to determine growth delay and regrowth, respectively, after drug treatment requires long-term culturing (> or =14 d).

Choosing the right cell line for breast cancer research

Abstract: Breast cancer is a complex and heterogeneous disease. Gene expression profiling has contributed significantly to our understanding of this heterogeneity at a molecular level, refining taxonomy based on simple measures such as histological type, tumour grade, lymph node status and the presence of predictive markers like oestrogen receptor and human epidermal growth factor receptor 2 (HER2) to a more sophisticated classification comprising luminal A, luminal B, basal-like, HER2-positive and normal subgroups. In the laboratory, breast cancer is often modelled using established cell lines. In the present review we discuss some of the issues surrounding the use of breast cancer cell lines as experimental models, in light of these revised clinical classifications, and put forward suggestions for improving their use in translational breast cancer research.

Animal cell cycles and their control

Abstract: THE ONSET OF M PHASE IN ANIMAL CELLS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ... . . . . . . . . . 444 Maturation-Promoting Factor and Growth-Associated Kinase 444 p34cdc2: the Catalytic Subunit of MPF .. . . ... . . . . . . . . . . . . . . . . . . . . . .. . . . ... . . . . . . . . . . . . . . . . . . . 445 Cyclin B: the Second Essential Component of MPF. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 446 Regulation of p34cdc2 Activity by Phosphorylation . . . . ...... . ..... . . . . . . . . . . . . .. . . . . . . . .. . . 447 Substrates of p34cdc2 and their Significance for M Phase Events . . . . . . . . . . . . . . . . . . . . . . . 450 Dependence of Mitosis on Completion of DNA Replication . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 453 Meiotic Maturation and Cytostatic Factor. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 455 Cell Cycle Roles for Phosphoprotein Phosphatases . . . . . . . . . . . . . .. . . . . . . . . . . . 456 CONTROL OF THE ANIMAL CELL CYCLE IN G I . . .... . . . . . . .. . . . ....... . . . . . . . . . . . .. . . 457 Growth Factor Requirements for Commitment of Fibroblasts to S Phase Entry 458 Protein Synthesis Requirements for Commitment of Fibroblasts to S Phase Entry. . . . . . . . . . . . . . ......... . . . . .. . . . . . . . . . . . . 460 The Search for an S Phase-Promoting Factor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 460 Growth Control and Cell Cycle Control . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 463 PROSPECTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 464