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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

Copyright © 2011 by Republic Polytechnic, Singapore

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.

Copyright © 2011 by Republic Polytechnic, Singapore

FROM WHERE ARE CELLS LINES OBTAINED?

Cells lines

Copyright © 2011 by Republic Polytechnic, Singapore

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

Copyright © 2011 by Republic Polytechnic, Singapore

HOW ARE CELL LINES DIFFERENT?-DIFFERENT MORPHOLOGIES OF MAMMALIAN CELL IN CULTURE

Copyright © 2011 by Republic Polytechnic, Singapore

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 ?

Copyright © 2011 by Republic Polytechnic, Singapore

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

Copyright © 2011 by Republic Polytechnic, Singapore

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

Copyright © 2011 by Republic Polytechnic, Singapore

Differences between growth requirements of adherent and suspension cell lines?

HOW ARE CELL LINES MAINTAINED ?

Copyright © 2011 by Republic Polytechnic, Singapore

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.

Copyright © 2011 by Republic Polytechnic, Singapore

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