2017 Lab 9

From CellBiology

Introduction

In the lab I will be covering:

  1. The Signaling 2 lecture, which should be considered part of the examinable theory
  2. Completing the group project work
  3. Finishing the T/C practical information.
  4. 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.



Projects

2017 Project Topics - TBD.


  • 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

  1. The key points relating to the topic that your group allocated are clearly described.
  2. The choice of content, headings and sub-headings, diagrams, tables, graphs show a good understanding of the topic area.
  3. Content is correctly cited and referenced.
  4. 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.
  5. Evidence of significant research relating to basic and applied sciences that goes beyond the formal teaching activities.
  6. Relates the topic and content of the Wiki entry to learning aims of cell biology.
  7. 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.
  8. Evaluates own performance and that of group peers to give a rounded summary of this wiki process in terms of group effort and achievement.
  9. 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.
  10. Develops and edits the wiki entries in accordance with the above guidelines.


Cell Growth

Cell Media

Media-bottle-icon.jpg

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.


Tissue culture flasks

Different Cell Metabolic requirements

Serum

  • 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).

Serum-Free Media

  • Stem cell and human therapeutic applications.
  • Allow growth in a fully-defined media.
  • No batch to batch sera variations.
  • Eliminate cross-species contamination/interactions.

Antibiotics

Links: Sigma - Cell Culture Tested Antibiotics

Typical 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

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

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

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.

Cell Analysis

Cells can be analysed in many different experimental techniques

  • Living or Fixed
  • Microscopically
    • Time lapse
    • Confocal microscopy
    • Immunohistochemistry
    • In Situ hybridization
  • Biochemically
    • DNA (Southern)
    • mRNA (Northern)
    • Protein (Western)
    • Microarray
  • Flow cytometry

Practical Methods Video

Journal of Visualized Experiments

Stem Cell Practical - Exercise

2017 Lab 11

PDFs of 3 suggested primary research articles on stem cell research published any time from 2012 till now by the end of this week.


As part of the assessment for this course, your group will give a 15 minutes journal club presentation during the Lab on 30 May. For this you will have to discuss a recent (published after 2012) 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 19 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



2017 Course Content

Moodle

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 | 2017 Revision

2017 Laboratories: Introduction to Lab | Fixation and Staining |


2017 Projects: Group 1 - Delta | Group 2 - Duct | Group 3 - Beta | Group 4 - Alpha

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