APR January/February 2022 - 31

« DRUG DEVELOPMENT
mandatory product (Quality Control) testing
so that their existence will be recognized and
the cells discarded before use. There are some
potential, and possibly some as yet unknown
pathogens, which may be able to incorporate
themselves into cells and establish persistent
yet non-evident infection. These infective
agents may originate from the donor cells
themselves, contamination at the time
of harvesting, or during the propagation
process.
There are two options envisaged/employed
in obtaining starting cells as an autologous
starting cell line to process and produce the
medicinal products;
1. The use of patient own cells as
starting material, Figure 3.
2. The use of compatible (other than
the patient) cells as a starting
material, Figure 4.
What is Transfection
and How to Transfect
Cells
Transfection is the process of deliberately
introducing naked or purified nucleic
acids into eukaryotic cells. It includes the
introduction of DNA, RNA, or proteins into
eukaryotic cells and is used in research to study
and modulate gene expression. Transfection
techniques serve as an analytical tool that
facilitates
the
characterization
of genetic
functions, protein synthesis, cell growth and
development. Transfection assays enable the
advancement of cellular research to enhance
drug discovery strategies. Strategies such as
viral transfection or viral transduction, utilize,
for example, lentiviral (see Table 1) particles
to insert foreign material into eukaryotic cells.
Types of Transfection
There are a wide range of transfection
methods being utilized including physical,
chemical and biological techniques. These
techniques generally involve the use of
transient or stable transfection methods to
incorporate nucleic acids into cells.
Transient transfection techniques involve
the introduction of DNA into cells, but in this
method, the DNA does not integrate with
the cellular chromosomes. This technique
facilitates
high transfection efficiencies
and the gene transcripts can be analyzed
after a period of one to four
days. For
large-scale transient gene expression (TGE)
in mammalian cell cultures, transfection
vehicles such as polyethylenimine (PEI) and
calcium phosphate (CaPi) can be used. Largescale
TGE methods have also been developed
using Chinese Hamster Ovary (CHO) cells in
the absence of serum.1
Some of the commonly used transfection
techniques include calcium phosphate
precipitation, lipofection, electroporation,
perfection is all-around_
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APR January/February 2022

Table of Contents for the Digital Edition of APR January/February 2022
