2010 Lab 9

Introduction

Biohood

The previous laboratory introduced cell culture, cells and practical considerations. This laboratory will look at the more "hands on" techniques required to carry out tissue culture.

Practical Methods Video

• Sigma - Aseptic Techniques

• Sigma - The Micropipette

• Journal of Visualized Experiments
  • Counting and Determining the Viability of Cultured Cells
  • Trypsinizing and Subculturing Mammalian Cells
  • Cell Capture Using a Microfluidic Device

Practical Methods

• General Guide For Cryogenically Storing Animal Cell Cultures (Corning)
  • This 10-page PDF Corning Technical Guide examines both the theoretical and practical aspects of cryogenic preservation and reviews key strategies for managing a cell repository.

Cell Lines

C2C12 Myoblast

• Mouse (Mus musculus) muscle cell line

• Original Reference Blau HM, et al. Plasticity of the differentiated state. Science 230: 758-766, 1985. PubMed: 2414846

• ATCC ATCC C2C12 | ATCC C2C12 images

• Blau Lab Protocol Blau Lab - C2 Mouse Myoblast Culture

PC-12 Pheochromocytoma

• Rat (Rattus norvegicus), adrenal gland cancer

• Original Reference Greene LA, Tischler AS. Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc. Natl. Acad. Sci. USA 73: 2424-2428, 1976. PubMed: 1065897

• ATCC ATCC PC12 | ATCC PC12 images

3T3 Fibroblast

• Mouse, Swiss albino (Mus musculus)

• Original Reference Todaro GJ, Green H. Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. J. Cell Biol. 17: 299-313, 1963. PubMed: 13985244

• ATCC ATCC 3T3 | ATCC 3T3 images

HeLa Epithelial

• Human (Homo sapiens) cervical adenocarcinoma

• HeLa cells were named for Henrietta Lacks, who died in 1952 from cervical adenocarcinoma and her physician Margaret Gey then began working with these cells used throughout the world for medical research.

• ATCC ATCC HeLa | ATCC HeLa images

References

Textbooks

Molecular Biology of the Cell

Alberts, Bruce; Johnson, Alexander; Lewis, Julian; Raff, Martin; Roberts, Keith; Walter, Peter New York and London: Garland Science; c2002

Growth Factor Model Image: MBoC Ch18 Fig 45 MCB Figure 6-5. Stages in the establishment of a cell culture. Image: MBoC Ch18 Fig 46/47

• Some Landmarks in the Development of Tissue and Cell Culture
• Cells Can Be Grown in a Culture Dish
• Serum-free, Chemically Defined Media Permit Identification of Specific Growth Factors

Molecular Cell Biology

Lodish, Harvey; Berk, Arnold; Zipursky, S. Lawrence; Matsudaira, Paul; Baltimore, David; Darnell, James E. New York: W. H. Freeman & Co.; c1999

• Growth of Animal Cells in Culture
• Figure 6-5. Stages in the establishment of a cell culture

The Cell- A Molecular Approach

Cooper, Geoffrey M. Sunderland (MA): Sinauer Associates, Inc.; c2000

• Growth of Animal Cells in Culture

Search Online Textbooks

• "cell+culture" Bookshelf

PubMed

• PubMed is a service of the U.S. National Library of Medicine that includes over 18 million citations from MEDLINE and other life science journals for biomedical articles back to 1948. PubMed includes links to full text articles and other related resources. PubMed

• PubMed Central (PMC) is a free digital archive of biomedical and life sciences journal literature at the U.S. National Institutes of Health (NIH) in the National Library of Medicine (NLM) allowing all users free access to the material in PubMed Central. PMC

• Online Mendelian Inheritance in Man (OMIM) is a comprehensive compendium of human genes and genetic phenotypes. The full-text, referenced overviews in OMIM contain information on all known mendelian disorders and over 12,000 genes. OMIM

• Entrez is the integrated, text-based search and retrieval system used at NCBI for the major databases, including PubMed, Nucleotide and Protein Sequences, Protein Structures, Complete Genomes, Taxonomy, and others Entrez

Search Pubmed

• "tissue culture" Entrez all databases
• "cell culture" Entrez all databases

Search PLoS

• PLoS One ribosome

Reviews

• PMID: 18365235

Books

• Animal Cell Culture: A Practical Approach ed R.I Freshney

2010 Course Content

Lectures: Cell Biology Introduction | Cells Eukaryotes and Prokaryotes | Cell Membranes and Compartments | Cell Nucleus | Cell Export - Exocytosis | Cell Import - Endocytosis | Cell Mitochondria | Cell Junctions | Cytoskeleton Introduction | Cytoskeleton 1 Intermediate Filaments | Cytoskeleton 2 Microtubules | Cytoskeleton 3 Microfilaments | Extracellular Matrix 1 | Extracellular Matrix 2 | Cell Cycle | Cell Division | Cell Death 1 | Cell Death 2 | Signal 1 | Signal 2 | Stem Cells 1 | Stem Cells 2 | Development | Revision

Laboratories: Introduction to Lab | Microscopy Methods | Preparation/Fixation | Immunochemistry | Cell Knockout Methods | Cytoskeleton Exercise | Confocal Microscopy | Microarray Visit | Tissue Culture 1 | Tissue Culture 2 | Stem Cells Lab | Stem Cells Analysis

Dr Mark Hill 2015, UNSW Cell Biology - UNSW CRICOS Provider Code No. 00098G