Our
concept of laboratory design and cell culture continues to evolve. What many
don’t comprehend is how the pioneers in the field of cell culture recognized
the impact of the laboratory’s design on the success of their efforts. Regardless,
of the exact nature of cell culture: embryo culture, cell culture for virus
propagation, production of recombinant products, all cell culture techniques
must provide a suitable environment for both the culture and the personnel
working in the facility.
Cell
culture started in the late 19th century with cells and tissues being isolated
and grown in a wide variety of biological media such as vitreous humor, clotted
lymph and plasma. Early efforts often used a “hanging drop” method
where a small section of tissues was inserted in a clot of chick serum and
plasma. The “incubators’ of the time were modified poultry incubators.
By the early 20th century, Alexis Carrel had developed methods for continuous
culture of cells. Then with Earle’s work “chemical defined” entered
the field. This resulted in culture methods with individual cells in
suspension or as a monolayer growing on the surface of the containing vessel.
By today’s standards, the facilities were primitive. Sterility was the gold
standard, since antibiotics were not available.
By
the midpoint of the 20th century, specific laboratory designs were available in
the literature. If we went back to these facilities we would recognize
many of the goals but not the methods.
Raymond
Parker in the 3rd edition of “Methods of Tissue Cultures” published in 1961
describes some of these historical and then “state of the art” laboratories.
Alexis Carrel (1912 Nobel Laureate), at the Rockefeller Institute, tried to
reduce glare and eyestrain by using natural lighting combined with walls painted
gray and black. Hospital white, then the common practice, was replaced by
Carrel’s black gowns and drapes. To settle dust before HEPA filtration, a
series of water mist spray heads were built in the ceiling. They would be
used after laboratory work had ceased for the evening. The mist would be
allowed to settle the dust overnight. Utilities were built in to the walls
to eliminate areas for the accumulation of dust. Carrel’s lab had a
series of rooms transitioning from cleaning and sterilizing to the more tightly
controlled rooms used for tissue preparation and culture.
Parker’s
laboratory at the University of Toronto used a series of interconnecting rooms
to provide isolation. “Forced filtered ventilation” was used to remove dust
and pressurize the laboratory. He called for terminal particulate
filtration at the end of air supply ducts. He specified that the only
outlet for escaping air would be the entry door. He recognized the need
for the sealing of all outlets and wall penetrations.
While
particulate filtration was improved by the fifties and sixties, germicidal lamps
were required to operate continuously. For temperature control ceiling
mounted cooling coils were used. Heat was supplied by radiators built into
walls. Recalling some of the early days of embryo culture, Parker’s
cultures were incubated in constant temperature rooms set at 37 degrees C.
The rooms were large enough to provide room for staff to perform microscopic
work and manipulations. It must have brought a whole new meaning to
working over a hot microscope, though some IVF laboratories operated this way in
the 80s.
Cleaning
in these early laboratories was a nightmare. Glassware was the only material and
preparation for culture included washing in strong acids and alkali to prepare
the surfaces for cell attachment. Sterilization was done by a combination
of dry heat and autoclaving. Ultrasound cleaning tanks were considered a
major step forward in aiding the cleaning process. Clearly, our use of
prepackaged plastic ware has eliminated this major headache of pioneers in cell
culture.
Please
note that Galileo Research Laboratories, its staff, or any of its associated
members are in no way responsible for or necessarily agree with the opinions
presented here.
