This document provides an overview of animal cell and tissue culture. It begins by defining animal cell culture as dealing with controlling and modulating cellular function, recognizing cells as independent organisms. Examples are given of transforming cells into engineered systems to express specific biological functions in vitro. Considerations for animal cell culture models include choosing appropriate cell types and designing systems that mimic human physiology. Requirements for setting up an animal cell culture like media, incubators, and safety equipment are also outlined. Specific techniques are discussed such as obtaining skin cells and culturing types of liver cells. Requirements for the next class on differences between animal and cell experiments are assigned.

1. BTE 4223
DR. MUNIRA SHAHBUDDIN
KULLIYYAH OF ENGINEERING
IIUM
LECTURE I
ENGINEERING ASPECT OF
ANIMAL CELL AND TISSUE
CULTURE

2. INTRODUCTION
• WHAT IS ACTUALLY ANIMAL CELL CULTURE?
The main subject of animal cell technology is to deal with control and
modulation of cellular function.
Conventional cell biology regards cells and tissues as constituent of
animal body.
The analyses of differentiated cellular function give the basic
information needed to understand composite functions of organs and
animals which consist of organized assemblies of various kinds of cells

3. • Animal cells technology
recognizes an animal cells as an
independent living organism

4. EXAMPLES OF TRANSFORMATION FROM
CELLS INTO A SYSTEM

5. • Animal cell technology is very interested in focusing on the
construction and functional regulation of engineered cells which
are reconstituted so as to express their specific biological
function in vitro.
• Animal cell culture technology is a new scientific discipline
closely associated with the further development of
biotechnological industrial field.

6. • The research on bio-functional regulation of engineered cells
will lead to a much more work and clear understanding of
cascades of an in vivo physiological events.
• Biological function of engineered cells can be controlled and
modulated by various means which are categorized into three:
genetic, physiological and physical means.

7. CHOOSING THE APPROPRIATE ANIMAL CELL
CULTURE MODEL
• Why opt for animal cell culture? What is so wrong
with animal sacrifices?
• Cost analysis and benefit.
• The use of animal cells will reduce the cost for
animal care and housing, less time consuming and
much cleaner.
• Effective and easy.
• Rules and regulation of economic block and
countries

8. • The test for cosmetics and household products should not be so cruel
but rather simple, easy, rapid and effective.
Left: Inflammatory test on rabbit’s eye and Right: on human tissue engineered skin

9. CONSIDERATIONS FOR ANIMAL CELL CULTURE
• The vision or model of an animal cell culture system as difficult,
inefficient and uneconomic is a result of the perception that it
requires expensive media, easily contaminated and difficulties for
scaling-up process.

10. • The implementation of Good Manufacturing Practices has led to the
almost complete elimination of contamination.
• There are considerations that should be taken while doing cell
culture: oxygen supply and demand, oxygen delivery system, air flow
rates, bubble diameter, oxygen transfer rate, bubble residences,
cellular physiologies, oxygen solubility, pH, and etcetera.

11. WHY CONSIDER?
1. Think of a system that mimic human
physiology, what do you require?
2. What will be the effect of the system
microenvironment to cellular physiology
and biological effects?
3. What is cellular microenvironment?
Designing an experiment or model encourage
us to work outside the conventional practices.
In general it provides us with the basis for
further investigations to improve our
understanding on cellular biology and system.

12. REQUIREMENTS FOR ANIMAL CELL CULTURE
• Cell culture media
• Incubator
• Laminar hood
• Laboratory apparatus – pipettes, petri dishes, beaker etc.
• Disinfectant – bleach and 70% ethanol
• Cryopreservation medium

14. An easy way to obtain epidermal skin…….
• When cells from adult foreskin epidermis were cultured
in vitro for 7–10 days, small fusiform cells appeared (P0
7d), while other cell types almost completely
disappeared. With prolonged incubation, these small
fusiform cells continued to increased in number (P0 12d
and P0 15d). (B): Before passage 10, most cells were
short and spindle-shaped (P2 2d). Within the first 30
passages, cells generally formed a single layer within
1.5–3 days after replating (P2 3d). (C–E): After passage
10, most cells had short spindle or regular spindle
morphologies, with two to three processes projecting
from the soma (C, round figure on the left, see arrow),
while a minority had multiple processes (C, round figure
on the right, see arrow). In some specimens, most of the
hEMSCPCs had short processes, while only a minority
had long and slim processes (D, round figure on the left,
see arrow). In other specimens, however, most of the
hEMSCPCs had long and slim processes. (F): When
hEMSCPCs formed a monolayer, they took on a swirling
arrangement resembling that of mesenchymal stem
cells.
Reference:
http://www.nature.com/articles/srep01933

15. Reference to the paper: Organotypic liver culture models: Meeting current challenges in toxicity
testing. LeCluyse EL, Witek RP, Andersen ME, Powers MJ - (2012)
http://openi.nlm.nih.gov/detailedresult.php?img=3423873_btxc42-501-
f8&query=null&req=4&npos=-1

16. fig5: Major nonparenchymal cell types of the liver. Top row: CD-31 staining of
liver sinusoidal endothelial cells (LSEC) lining vascular walls of whole liver (A),
scanning electron micrograph of the endothelial lining of the liver sinusoids
showing extensive patches of fenestrae (arrows) (B), and primary LSEC
showing typical morphology in vitro (C). Middle row: HSC (GFAP) in normal
liver (D), myoflbroblastic HSC ((iSMA) in flbrotic liver (E), and isolated qHSC
showing storage of vitamin A as bright “floating” vesicles within the cell body.
Upon activation or injury the HSC undergo extensive morphological and
biochemical changes, which include the synthesis, secretion and restructuring
of ECM molecules. Bottom row: Kupffer cells (KC) showing their dynamic
morphology (G), their identification with CD68 showing extended projections
on the cell bodies used for contact with other cells (H), and a magnified view
showing KC loaded with vesicles containing cytokines and other secretory
factors (I).
fig8: Hepatic cell culture model systems represented in “Advanced
organotypic culture technologies” (A) Perfusion array liver system
(PEARL) (Lee et al., 2007), (B) bioengineered micropatterned liver
platform (Khetani and Bhatia, 2008), (C) biochip dynamic flow system
(Chao et al., 2009; Novik et al., 2010; Maguire et al., 2009), (D) 3-D
liver tissue culture scaffold (Sibanda et al., 1993; Sibanda et al., 1994;
Naughton et al., 1994), and (E) 3-D scaffolds with dynamic flow
(Sivaraman et al., 2005; Domansky et al., 2010).

17. • For the next class, I would like you to read on the differences between
animal and animal cell experimentation for discussion.
• Everyone is required to read on animal cells and tissue laboratory
manuals and regulations.