3.i. LECTURE NOTE ON CELL CULTURE

 

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DETAILS OF THE MODULE AT LUTON

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Handbook Description & Rationale

This module is designed to provide you with a basic training in cell culture techniques widely used in modern research laboratories and bio-industries. Although you will be trained to grow animal cells the basic techniques are not only directly applicable to human and other mammalian cells but also, more generally, to other cells, such as insect, or plant cells. The module is particularly useful if you wish to undertake a project in cell or tissue culture and it is assessed purely by the course work you produce throughout the semester.

Aims & Objectives

Time Allowed 7-12 hrs lectures, tutorials or seminars, 20-30 hrs practical,

40-70 hrs directed reading, 30 hrs assessment

Teaching Team TO BE ANNOUNCED

Prerequisites BHS01-1 Cell Biology & Genetics or

BHS05-1 Introduction To Biotechnology (or equivalent learning)

Learning Outcomes

At the end of the module the student should be able to:

Learning Strategies

This module is heavily practical based and this is reflected in the teaching and learning strategy. The background to the subject area will be delivered using a mixture of lectures, videos and directed reading and study. A mixture of videos and demonstrations will be used to provide the students with the basic training in the necessary techniques. Although there will be initial instruction in the basic techniques and monitoring of progress the student will be encouraged to plan, organise and conduct the majority of the day to day work themselves, in consultation with the supervisory team. Students will keep a diary of their practical work undertaken, i.e. procedures carried out, growth of cells, problems met and solved etc.

Towards the end of the module there will an assessment of the students practical skills in cell culture. There may also be assessments of practicals during the module to give feedback to each student on their progress, but such assessments will not count toward the final grade, i.e. they will just be informative.

The progress of each student will also be monitored throughout the course through their interactions with the members of the teaching team and through regular inspection of the student’s diary by members of the teaching team.

LECTURES/TUTORIALS

Thursdays C303 9.00 – 10.00 am

PRACTICALS (see comments on attendance below)

Weeks 3- 6/7 C315

Weeks 7/8 – 12/13 C205 (Specialised cell culture laboratory)

ASSESSMENT DETAILS (DEGREE & HND)

Assessment 1 (50%): Practical Report

Composed of: (i) A report introducing, summarising and discussing all the practical work undertaken; submitted week 13 (25%).

(ii). Diary of laboratory work, submitted week 13 (25%).

Attendance Record. You must:

 

Relevant Skills

Social Development & Interaction: You should be able to:

Planning & Problem Solving: You should be able to:

Assessment 2 (50%): Practical Skills assessment

This will be assessment of student’s practical skills, undertaken as described as an end of module practical assessment in weeks 12/13

Relevant Skills

Content

 

OUTLINE TEACHING SCHEDULE

Weeks 1 to 7/8:

Lectures: Introduction to tissue culture: set of formal lectures, supported by videos of appropriate techniques

Practicals: Introduction to tissue culture techniques including: demonstration & learning of basic laboratory and tissue culture skills including: safety in handling cell lines; how to handle cells; how to count cells, how to assess their viability, etc.

Week 8/9 to 14

Lectures: There will no formal lectures for most of this period, but students will have the opportunity to discuss particular problems in the actual laboratory sessions and specific tutorials will be arranged to cover the assessment points and others if needed/requested by the students.

Practicals: Series of demonstrations and practical sessions on basic cell culture skills, including; use of equipment and appropriate sterile techniques; how to culture (grow) cells; how to sub-culture (passage) cells; how to monitor viability and success of culture techniques; how to store cells, leading up to the assessment of student’s practical cell culture skills.

 

PRACTICAL DETAILS

NOTE:

**NOTE: Students will also get a chance to complete their cell dilution and counting experiments on Tuesdays, in the anti-chamber to the cell culture lab in C205. Students need to arrange a time to do this (on a Tuesday) with the relevant demonstrator..

READING MATERIAL & VIDEOS HELD IN THE LIBRARY AT LUTON

Core Text

  1. Animal cell culture. Sara J. Morgan, David C. Darling. Oxford : BIOS Scientific in association with the Biochemical Society, 1993. - xiii, 162p ; pbk. - 1-872748-16-3 (£18.95 Jan. 1999 price)
  2. NOTE: The preferred core text is out of print this year (2001). Reference No. 2 (below) has been given to the bookshop as the recommended text for this year. However, there are other books, equally as good as the preferred text (No. 1 above) in the library. Students may like to assess these themselves, as compared to No. 1 above and order the one that they preferred. Note, student planning to take the third year module, Applications of Immunology, should try and get the recommended text as it has some parts that will be useful for this module too.

    Other Reading

  3. Animal cell culture. A Practical Approach. Ed I. Freshney. The Practical Approach Series. Eds Rickwood D. and Hames B.D. IRL Press, Oxford University Press. New York. 1992. 2nd edition. [ISBN: 0 19 963213 8].
  4. Introduction to Light Microscopy. S. Bradbury, B. Bracegirdle. Oxford: Bios Scientific, 1998 [ISBN: 1-859961-21-5]
  5. Alison, M.R. Assessing cellular proliferation: what's worth measuring? Offprint from Human & Experimental Toxicology 14 1995. - x-34-052667-9 and - x-34-081116-0
  6. Animal cell culture methods. G.D. Wasley, John W. May. Oxford : Blackwell Scientific, 1970. - vi,194p : ill, 1form ; 23cm. bibl p179. - 0-632-04240-0*
  7. Cell culture: Labfax. Ed M. Butler and M. Dawson. Oxford : BIOS Scientific Publishers [with] Blackwell Scientific Publishers, 1992. - xix, 247p : ill. - (Labfax Series) For use in laboratories. - Includes index. - 1-872748-10-4
  8. Cell culture. Ed William B. Jakoby, Ira H. Pastan. San Diego /etc./ : Academic Press, 1979. - xxi, 642p. - (Methods in enzymology ; 58). - 0-12-181776-8
  9. Methods in cell biology. - Volume 21: Normal human tissue and cell culture. Ed Curtis C. Harris, Benjamin F. Trump, Gary D. Stoner. - B: Endocrine, urogenital, and gastrointestinal systems. New York : Academic Press. 1980. - xvii, 512p : ill. Prepared under the auspices of the American Society for Cell Biology. - 0-12-564140-0
  10. Methods in enzymology Vol 58. Cell culture. Academic Press. - x-34-004851-3
  11. Methods in molecular biology. - Vol.5: Animal cell culture. Ed by Jeffrey W. Pollard and John M. Walker. Cifton, N.J. : Humana Press, 1990. - 0-89603-150-0
  12. Jones, Michael Kenyon. The application of a modified cell perifusion system to the study of adenosine metabolism in isolated immobolised adipocytes. Luton : LCHE, 1992. - 345 leaves. M.Phil thesis (CNAA), Luton College of Higher Education, 1992. - x-34-070507-7
  13. Practical tissue culture applications. Ed Karl Maramorosch and Hiroyuki Hirumi. New York : Academic Press, 1979. - xv,426p : ill ; 23cm. 'This book contains the proceedings of a conference held at the International Laboratory for Research on Animal Diseases in Nairobi, Kenya, August 24-29, 1978' - half title page verso. – Includes bibliographies and index. - 0-12-470285-6
  14. Tissue and cell culture (by) A.M. Whitaker. London : Baillière Tindall, 1972. - viii,119,(2)p(2fold) : ill ; 22cm. - (Laboratory monographs) Bibl.p.107-110. - Index. - 0-7020-0413-8
  15. The isolation and culture of rat hepatic cells. Offprint from INVITTOX Protocol 7 June 1989. - x-34-081331-7

Relevant Videos in the Library
(please click here to download a list of videos in word.doc format)

601. Basic Sterile Cell Culture: Sterile cell culture techniques are shown by documenting actual experiments. Maintenance and passage of cultures are documented. The investigators use techniques such as disinfecting instruments and work areas, aseptically pipetting cell cultures, using a laminar flow hood and other subtle methods for ensuring sterile viable cultures. Evidences and causes of contamination are described. 17 mill. ©1992. $59.00 (in library from May 2001)

602. Microscopically Characterising Cells: Video microscope images are used to characterize commonly used cells. Cells of different morphologies, shapes, sizes, growth patterns and organelle definitions are seen through a phase contrast microscope. Different percentage confluency of cell cultures and identifying features of dead or viable cells are shown. 16 min. ©1992. $59.00 (in library from May 2001)

603. Quantifying Viable Cells: The proper method for quantifying viable cells using a hemocytometer is documented in an actual experiment. Viable cells are distinguished from unhealthy cells with and without using trypan blue exclusion. Other methods for quantifying cells using a

Coulter Counter and fluorescent activated cell sorter are illustrated briefly. 12 min. ©1992. $49.00 (in library from May 2001)

604. Cryopreserving Cultured Cells: An important part of cell culture is the ability to cryopreserve or freeze cells as a method of storing them, which is the subject of this video. The method and principles of cryopreserving cells using DMSO with minimal loss of viability are shown in an actual experiment. The investigator uses glass vials or ampules to effectively store frozen cells in liquid nitrogen and compares their use with plastic vials. 22 min. ©1992. $62.00 (in library from May 2001)

605. Trypsin-Treatin Attached Cells: Many commonly used cell lines require a solid matrix on which to grow, such as tissue culture plastics. Once they take up all the space on the surface of the plastic, they need to be removed with trypsin-treatment and passed as new cultures, which is the subject of ntis video. 7 min. © 1992. $39.00 (in library from May 2001)

RELEVANT WEB SITES (please see other pages on this site for other links)

http://www.research.umbc.edu/~jwolf/method5.htm

http://www.jswresearch.com/neuronaltissueculture.html

http://hope.21cn.com/lab/pages/cell%20culture.htm