2016 Lab 9
- 1 Introduction
- 2 Cell Signaling 2
- 3 Projects
- 4 Tissue Culture Practical 2
- 5 Stem Cell Practical - Exercise
- 6 2016 Course Content
In the lab I will be covering:
- The Signaling 2 lecture, which should be considered part of the examinable theory
- Completing the group project work
- Finishing the T/C practical information.
- Preparing the students for the stem cell presentations.
Also Dr Fath today will give an introduction to a second semester course, ANAT 3212 Microscopy in Research. See the attached course outline from 2015 ANAT 3212 Microscopy in Research.
Cell Signaling 2
This lecture is the second in a series of lectures on cell signaling. The first lecture was a general introduction to the different signaling mechanisms and steroid pathways. The following lecture looks in detail at two specific examples of signaling pathways: G Protein-Coupled Signal and Receptor Tyrosine Kinases. Note that you have already been introduced to signaling in many of your other lecture topics and the lectures on stem cells and development will further discuss cell signaling events.
- Group projects are due for completion this week.
- Project pages will then be locked for my assessment.
- I will be pasting my assessment information for each project at the top of tha discussion page. Please note that this is an ongoing process, and individual student marks are not displayed in the assessment section.
- Project Discussion pages will remain unlocked, so that you can update any group discussion information. If you intend to cut and paste from another source, please replace any identifying (name) information with student IDs. Simply search and replace in a word document before pasting on discussion page.
Group Assessment Criteria
- The key points relating to the topic that your group allocated are clearly described.
- The choice of content, headings and sub-headings, diagrams, tables, graphs show a good understanding of the topic area.
- Content is correctly cited and referenced.
- The wiki has an element of teaching at a peer level using the student's own innovative diagrams, tables or figures and/or using interesting examples or explanations.
- Evidence of significant research relating to basic and applied sciences that goes beyond the formal teaching activities.
- Relates the topic and content of the Wiki entry to learning aims of cell biology.
- Clearly reflects on editing/feedback from group peers and articulates how the Wiki could be improved (or not) based on peer comments/feedback. Demonstrates an ability to review own work when criticised in an open edited wiki format. Reflects on what was learned from the process of editing a peer's wiki.
- Evaluates own performance and that of group peers to give a rounded summary of this wiki process in terms of group effort and achievement.
- The content of the wiki should demonstrate to the reader that your group has researched adequately on this topic and covered the key areas necessary to inform your peers in their learning.
- Develops and edits the wiki entries in accordance with the above guidelines.
Tissue Culture Practical 2
Basic Constituents of media, inorganic salts, buffering systems, carbohydrates, vitamins, proteins/peptides, fatty acids/lipids, trace elements.
- cells require pH conditions in the range 7.2 - 7.4 and close control of pH.
- phenol red acts as a pH indicator, culture medium should be changed if the color turns yellow (acid) or purple (alkali).
- bicarbonate/CO2 buffering systems need to be maintained in incubator atmosphere of 5-10% CO2.
Different Cell Metabolic requirements
- Neuronal cells - High glucose (4500 mg/L) Dulbecco's Modified Eagle Medium (D-MEM) (1X) liquid (high glucose)
- Non-neuronal cells - low glucose (1000 mg/L) Dulbecco's Modified Eagle Medium (D-MEM) (1X) liquid (low glucose)
- Heat inactivation - involves heating at 56°C for 1 hour to inactivate complement components and prevent the occurrence of complement mediated lysis in cell cultures.
- Safety testing - different between countries: No safety testing required (USA/Canada, New Zealand, Finland and Denmark), Safety testing may be required (Australia, Mexico, Central America).
- Stem cell and human therapeutic applications.
- Allow growth in a fully-defined media.
- No batch to batch sera variations.
- Eliminate cross-species contamination/interactions.
Typical Cell Lines
- Mouse (Mus musculus) muscle cell line
- Original Reference Blau HM, et al. Plasticity of the differentiated state. Science 230: 758-766, 1985. PubMed: 2414846
- Blau Lab Protocol Blau Lab - C2 Mouse Myoblast Culture
- 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
- 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
- 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.
Cells can be analysed in many different experimental techniques
- Living or Fixed
- Time lapse
- Confocal microscopy
- In Situ hybridization
- DNA (Southern)
- mRNA (Northern)
- Protein (Western)
- Flow cytometry
Practical Methods Video
Journal of Visualized Experiments
- Journal of Visualized Experiments
Stem Cell Practical - Exercise
As part of the assessment for this course, your group will give a 15 minutes journal club presentation during the Lab on 26 May. For this you will have to discuss a recent (published after 2011) original research article on stem cell biology or technology, regenerative medicine, or stem cell therapy published in a high quality international research journal. Please note, reviews are not allowed.
Please search on PubMed for 2-3 research articles using search relevant terms, and send their PDFs to Annemiek (A.Beverdam@unsw.edu.au) by 5 pm on Wednesday 18 May. She will inform you which of the chosen articles are best suitable for presentation by Friday 20 May latest. Please note that the most exciting research articles are found in journals such as Nature, Science, Cell, Cell Stem Cell, etc. Please contact Annemiek in case you are at a loss, and she will help you find one.
During the presentation it works best if one student discusses the introduction, the second the results section, and the third the discussion section. Please note that one slide takes about 1 minute to talk through. So do not use more than 15 slides total. Please read through attached document for tips for how to prepare a good presentation. To make the Lab more interactive, I expect that each group will ask at least one question after each presentation.
You will receive a group mark based on presentation content, insight and comprehension, presentation and slide style, keeping within time, and contribution to discussion and questions.
Search PubMed: stem cell
2016 Course Content
Lectures: Cell Biology Introduction | Cells Eukaryotes and Prokaryotes | Cell Membranes and Compartments | Cell Nucleus | Cell Export - Exocytosis | Cell Import - Endocytosis | Cytoskeleton Introduction | Cytoskeleton - Microfilaments | Cytoskeleton - Microtubules | Cytoskeleton - Intermediate Filaments | Cell Mitochondria | Cell Junctions | 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 | 2016 Revision
Laboratories: Introduction to Lab | Microscopy Methods | Preparation/Fixation | Cell Knockout Methods | Cytoskeleton Exercise | Immunochemistry | Project Work | Confocal Microscopy | Tissue Culture | Stem Cells Lab | Microarray Visit
Dr Mark Hill 2015, UNSW Cell Biology - UNSW CRICOS Provider Code No. 00098G