Copyright © 2011 by Republic Polytechnic, Singapore ACTIVITY OWNER: RITU BHALLA (DR) APPROVED BY: VENTRIS D’SOUZA (DR) MODULE CHAIR: RITU BHALLA (DR) PROBLEM 3: WHO SELLS CELLS? A241 CELL CULTURE 6 TH PRESENTATION
Dec 03, 2014
Copyright © 2011 by Republic Polytechnic, Singapore
ACT IV ITY OWNER: R ITU BHALL A (DR)APPROVED BY: VENTR IS D ’SOUZA (DR)
MODULE CHA IR: R ITU BHALL A (DR)
PROBLEM 3: WHO SELLS CELLS?A241 CELL CULTURE6TH PRESENTATION
TODAY’S PROBLEM
• Your research team at the Skin Centre Research Centre has received a grant to study malignant melanoma
• You were asked to obtain suitable cell lines from a cell culture bank and maintain them
Copyright © 2011 by Republic Polytechnic, Singapore
WHAT DO YOU RECOGNIZE?
• Melanoma is related to skin cancer research• Cell lines can be used to study skin cancer• Cells may be obtained from a cell culture bank• There may be many different types of cell lines available
at the cell culture bank• It is necessary to choose cell lines that are the most
suitable for this project• One of the roles of a research assistant is to obtain cell
lines and maintain them • Cell lines need to be maintained in a lab.• There may be specific procedures required for
maintaining cellsCopyright © 2011 by Republic Polytechnic, Singapore
THE APPROACH
•What is malignant melanoma?•What is a cell line?•Where are cell lines obtained from? What is a cell culture bank? •How are cell lines different from each other? •How is an appropriate cell line selected?•Which cell line is suitable for the project?•What is required to maintain cell lines?•How are cell lines maintained?
Copyright © 2011 by Republic Polytechnic, Singapore
WHAT IS MALIGNANT MELANOMA ?
Malignant melanoma is a neoplasm (cancer) of melanocytes or of the cells that develop from
melanocytes
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in vitro cell culture assays are routinely used to conduct research studies on the causes and cure of melanoma.
WHAT IS A CELL LINE?A cell line is a propagated culture of specific cells after the initial
subculture, representing generations of a primary culture.
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FROM WHERE ARE CELLS LINES OBTAINED?
Cells lines
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Primary culture Isolation from tissue
Cell line culturesCell culture collections- ‘cell
banks’
HOW ARE CELL LINES DIFFERENT?
Cell lines differ in• their tissue of origin • cell morphologies and growth characteristics• life span (finite or continuous)• specific requirements for growth• genetic makeup
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HOW ARE CELL LINES DIFFERENT?-DIFFERENT MORPHOLOGIES OF MAMMALIAN CELL IN CULTURE
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Neuronal cells exist in different shapes and sizes
Fibroblastic (or fibroblast-like) cells are bipolar or multipolar, have elongated shapes, and grow attached to a substrate
Epithelial-like cells are polygonal in shape with more regular dimensions, and grow attached to a substrate in discrete patches
Lymphoblast-like cells are spherical in shape and usually grown in suspension without attaching to a surface
HOW IS AN APPROPRIATE CELL LINE SELECTED ?
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Characteristic Consideration
Species: organism of origin, disease or normal
Is the study human or non-human?
Finite or continuous: (Normal or transformed)
will cell line modification affect the study? What is duration of the study?
Bio safety level : I, II or III
what is the kind of facility available? experience in handling? nature of experiment?
Growth conditions & characteristics
What are your requirements with respect to growth rate, saturation density, cloning efficiency, and the ability to grow in suspension?
Functional characteristics
DNA profile, haploid/diploid? STR information,? is it suitable for drug testing?
Other criteria Is the cell line well-characterized? If using an abnormal cell line, is an equivalent normal cell line to use as a control available?
Depends o
n aim and sc
ope of the re
searc
h
study
WHY ARE THESE TWO CELL LINES SUITABLE FOR THE PROJECT?
Tumor cell line normal cell line
ATCC No. CRL-1619 CRL-1497
Designations A-375 [A375] CCD-34Sk
Organism Homo sapiens (human) Homo sapiens (human)
Morphology epithelial fibroblast
Source Organ: skinDisease: malignant melanoma
Organ: skinDisease: normal
Growth properties Adherent Adherent
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There are many possible cell lines that may be used for the project; the table below shows two of these cell lines
WHICH CELL LINE IS SUITABLE FOR THE PROJECT?
WHAT IS REQUIRED TO MAINTAIN CELL LINES?
• Sterile aseptic techniques• Optimal Growth conditions
• pH• Nutrients• Gas• Temperature• Surface?
• Growth maintenance
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Differences between growth requirements of adherent and suspension cell lines?
HOW ARE CELL LINES MAINTAINED ?
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WHAT YOU HAVE LEARNT …
• Understand what is a cell line.• Identify sources from where cell lines can be
obtained. • Differences between adherent and suspension
cells.• Considerations for selecting a cell line.• Requirements of media and procedures for
maintaining cell lines.
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End of Activity 3