06 Morphology of Caner Cells

The Biotechnology Education Company ®
EDVO-Kit #
990
Morphology of
Cancer Cells
Experiment Objective:
The experimental objective is to observe morphological
changes in cancer cells as compared to normal cells.
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EDVO-Kit # Morphology of Cancer Cells
990
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Table of Contents
Page
Experiment Components 3
Requirements 3
Background Information 5
Experimental Overview and General Instructions 7
Staining Procedure 8
Microscopic Observation 9
Study Questions 10
Instructor's Guide 11

General Information 12
Pre-Lab Preparations 13

Answers to Study Questions 14
Material Safety Data Sheets 15
All components are intended for

educational research only. They
are not to be used for diagnostic
or drug purposes, nor adminis-
tered to or consumed by humans
or animals.
EDVOTEK, The Biotechnology Education Company, and InstaStain are registered

trademarks of EDVOTEK, Inc.. Ready-to-Load and UltraSpec-Agarose are trademarks
of EDVOTEK, Inc.
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Morphology of Cancer Cells EDVO-Kit #
990
Experiment Components
• Ready-to-Stain two-spot slides containing cancer and normal cells

• Cell fixing agent
• Eosin stain
• Methylene blue
• Mounting medium
• Slide covers
• Transfer pipets
This experiment is designed
for 6 groups.
Store entire experiment

at room temperature.
Requirements
• Microscopes
• Forceps
• Distilled water
• Gloves
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information ready:
• Experiment number and title

• Kit lot number on box or tube
• Literature version number
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990
Background Information
Early studies on transformed malignant cells established that cancer-form-

ing cells exhibit unique and distinguishing features in comparison to their
normal cell counterparts. First, the cells are capable of sustained growth
in culture that was virtually immortal. Second, the cells overgrew in culture
forming a mass due to piling of cells. Third, solid tumor cells growing in
culture were less adherent to each other and the tissue culture substra-
tum. Fourth, the cells' morphologic appearance ranged from resembling
the normal cell from which they arose to an embryonic form. Finally, most
show an increase in number of nuclei, and distinct karyotype including
translocation, deletions, and chromosomal inversion.
Much of the current work is focused on how genetic alterations promote

the selections of cancer cells with aggressive behavior. Two classes of ge-
netic alterations occur in cancer including the gain of oncogenes and the
loss of tumor suppressors. Proto-oncogenes code for normal proteins in a
cell that are involved in accelerating growth. Oncogenes arise in cancer
cells when proto-oncogenes mutate or amplify leading to enhanced an
activity promoting cell growth. An analogy of how oncogenes act would
be that with the automobile whereby the gas pedal would be constantly
pushed down to increase the speed of a car. Tumor suppressors, other-
wise known as anti-oncogenes, are equally important in the development
of cancer. The normal function of tumor suppressor proteins are to restrict
cellular proliferation providing a protective function. When these genes
undergo mutagenesis or inactivation, cellular proliferation continues with-
out regulation, leading to cancer growth. Again the analogy of how tu-
mor suppressors act would be that with the automobile's brake pedal that
slows or regulates the rate of a cell's division. In cancer, the oncogene
takes over cell division and the altered or inactive suppressor no longer
has control over the process. In almost all cases the functional conse-
quence of these genetic changes is that cancer cells exhibit increased
growth, altered cell surface properties, loss of normal cellular morphology
and/or resistance to anti-cancer drugs, and increases in number of nuclei
and aberrant karyotype.
Although different cancer types, as well as cancer cells from a single tu-
mor mass may show a different genetic makeup and properties, the distin-
guishing feature of cancer cells is their loss of growth control. Cancer cells
show an abnormally high growth rate. This property of cancer cells is the
basis of how anti-cancer drugs selectively kill rapidly growing cancer cells
but do not harm slow growing normal cells with in a patient's body.
Cell growth involves an increase in cell mass and size which then triggers a
cell to divide. Thus cell growth and cell division are linked. Cell division in-
volves two recognizable coordinated events; the duplication of cell DNA
and physical division of the cells into two daughter cells. Because cancer
cells replicate so fast, they often show a multinucleated phenotype and
contain numerous nuclei. In addition, cancer cells often show an abnor-
mal variability in the size and shape of their nuclei. Pathologists often use
the abnormal nuclei observed in cancer cells to diagnose cancer.
Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/labora-
tory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the
written consent of EDVOTEK, Inc. Copyright © 2000, 2007, EDVOTEK, Inc., all rights reserved
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The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com
990
What contributes to the uncontrolled growth of cancer cells? Normal cells

show a property called contact inhibition, whereby cell growth ceases
under conditions of decreased nutrients, injury or cell crowding. Cancer
cells, however, have lost contact inhibition and continue to grow when
normally cells cease to grow. This increased growth leads to cancer cells
colonizing and destroying normal tissue. It is thought that defects in the

cell membrane of cancer cells interfere with the ability of these cells to
"sense" the cellular environment.
In addition to alterations in cell growth and loss of contact inhibition,

cancer cells show altered cell shape. Thus cancer cells look different from
their normal counterparts. Normal cells grow as ordered patterns as the
cell density increases. In contrast, cancer cells form chaotic masses. One
consequence of their altered morphology of cancer cells is that these
altered cell shapes contribute to increased cell movement seen in cancer
cells. A direct consequence of this defect leads to malignant cancer
cells migrating and spreading throughout the patient's body.
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990
Experiment Overview and General Instructions
Experiment Objective:
The experimental objective is to observe morphological changes in can-
cer cells as compared to normal cells.
LABORATORY SAFETY
The Experiment
1. Gloves and goggles should be worn routinely as good laboratory
practice.
2. Exercise extreme caution when working with equipment that is used in

conjunction with the heating and/or melting of reagents.
3. Do not mouth pipet reagents - use pipet pumps.
4. Exercise caution when using any electrical

equipment in the laboratory.
5. Always wash hands thoroughly with soap

and water after handling reagents or bio-
logical materials in the laboratory.
Laboratory notebook recordings:
Address and record the following in your laboratory notebook or on a

separate worksheet.
Before starting the Experiment:
• Write a hypothesis that reflects the experiment.

• Predict experimental outcomes.
During the Experiment:

• Record (draw) your observations, or photograph the results.
Following the Experiment:

• Formulate an explanation from the results.
• Determine what could be changed in the experiment if the ex-
periment were repeated.
• Write a hypothesis that would reflect this change.
EVT 009117AM
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Staining Procedure
1. Fix slides containing adherent cells by placing two to three drops of

fixing agent on each cell spot. Incubate 5 minutes at room tempera-
ture.
2. Air dry slides in hood.
3. Add one to two drops of blue stain (methylene blue) to each area of
the slide containing the cells.
The Experiment
4. Incubate the slide(s) for 7 minutes at room temperature.
5. Aspirate the methylene blue with a transfer pipet. Do not wash.
6. Add one drop of pink stain (eosin) to the same area. Incubate for 30
seconds at room temperature.
7. Rinse the slide(s) briefly with tap water. Gently tap or “wick” the slides
onto a paper towel to remove excess water. Mount the slide(s) by
placing one small drop of mounting medium onto the cells, then
carefully placing the coverslip(s) on top. View by bright field micros-
copy.
EVT 009117AM

990
Microscopic Observation
1. Locate cells with 10/ 20 X objective.
2. Describe the overall morphology of each slide section below.
3. Switch to high power. Ho do the number, shape, and size compare

between the different sections?
4. Classify each cell type as normal or cancerous.
The Experiment
Cell Low Power High Power Classification
Type Observations Observations
EVT 009117AM
10 EDVO-Kit # Morphology of Cancer Cells
990
Study Questions
1. What are the differences in cell density between the two slides?
2. Which cells have likely lost contact inhibition?
3. Which cells form clumps or piles of cells?

The Experiment
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06 Morphology of Caner Cells

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The Biotechnology Education Company ®

EDVOTEK, Inc. • 1-800-EDVOTEK • www.edvotek.com

Experimental Overview and General Instructions 7

Material Safety Data Sheets 15

All components are intended for

EDVOTEK, The Biotechnology Education Company, and InstaStain are registered

EDVOTEK - The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com

• Ready-to-Stain two-spot slides containing cancer and normal cells

Store entire experiment

EDVOTEK - The Biotechnology Education Company®

Visit our web site for information

• Experiment number and title

EDVOTEK - The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com

Early studies on transformed malignant cells established that cancer-form-

Much of the current work is focused on how genetic alterations promote

What contributes to the uncontrolled growth of cancer cells? Normal cells

colonizing and destroying normal tissue. It is thought that defects in the

In addition to alterations in cell growth and loss of contact inhibition,

Experiment Overview and General Instructions

2. Exercise extreme caution when working with equipment that is used in

3. Do not mouth pipet reagents - use pipet pumps.

4. Exercise caution when using any electrical

5. Always wash hands thoroughly with soap

Laboratory notebook recordings:

Address and record the following in your laboratory notebook or on a

Before starting the Experiment:

• Write a hypothesis that reflects the experiment.

During the Experiment:

Following the Experiment:

1. Fix slides containing adherent cells by placing two to three drops of

2. Air dry slides in hood.

4. Incubate the slide(s) for 7 minutes at room temperature.

5. Aspirate the methylene blue with a transfer pipet. Do not wash.

1. Locate cells with 10/ 20 X objective.

2. Describe the overall morphology of each slide section below.

3. Switch to high power. Ho do the number, shape, and size compare

4. Classify each cell type as normal or cancerous.

2. Which cells have likely lost contact inhibition?

3. Which cells form clumps or piles of cells?

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