Cell Growth Numbers
Country: United States
Date: September 2007
There is a limit to the number of some cells that can be
grown in culture; what are the conditions that cause these limitations?
If there was unlimited food and space on Earth, you could imagine that
our population would continue to grow forever, since our children could
always find a new place to live with food to eat. Earth, just like cells
growing in a culture, has a limited amount of space and resources. For
very similar reasons that our population can't continue to grow forever,
the population of cells growing in a culture will reach an upper limit
and eventually die off as nutrients in the media are depleted and toxic
cellular waste builds up (for example, some bacteria release acidic
metabolic byproducts which will lower the pH of the media solution).
In fact, the growth of bacteria in culture follows a predictable curve.
Initially, after a lag phase in which the cells are adjusting to their
new environment, nutrients are plentiful and the cells experience an
exponential growth phase. This is followed by a stationary phase where
environmental conditions are no longer as supportive and the rate of cell
death is similar to the rate of growth. Finally, when nutrients become
depleted and toxic waste products build up, cell deaths will exceed cell
divisions in what is known as a death phase.
As cells grow up, assuming you have an 'immortal' cell line (some cell lines
can't keep dividing indefinitely), there are physical limitations that
All cells need nutrients of some kind, and most industrial cultures need
oxygen as well. Large scale cell cultures are grown in large stirred tanks
called 'fermentors'. Fermentors typically have some kind of mixing paddle
along with inlets for air/oxygen and nutrients and outlets for off gases
Normally, you add a small number of cells to the tank (these are called
'seed' cells) along with food/nutrients and water (the food, nutrients, and
water are called the 'growth media', or 'media' for short). Hopefully, if
things go right, the cells will divide, and the culture will grow and become
more dense. As the culture grows, you have to keep adding air, and in many
cases, keep adding food as well.
As the culture gets more dense, it's harder to stir, and as a result, it's
harder to get the food and air into the cells. Cell near the air inlets
might be fine, but there might be places where the air and food don't reach,
and you can get 'dead zones' full of dead cells, or where the cells are
alive, but don't get enough food or air to grow and be healthy. This problem
is called "mass transfer limitation".
"Mass transfer" refers to how materials like food and air move around.
Imagine you are stirring food coloring in water. You give it a few stirs and
it's all evenly mixed; easy. However, what if you were mixing coloring into
paint? Paint stores have special machines to do this, but even with the
machines it takes a long time. If you tried to do it by hand it would take a
very long time (and give you sore arms too)! That's because the thicker
paint doesn't mix as easily as the thinner water. The same thing is true for
cells. As they grow, and the culture gets thicker, it gets harder to mix,
and harder to get nutrients into the cells.
Another consideration is that some cells actually make poisons that prevent
their own growth. Yeasts that make alcohol are a great example. When you
ferment beer or wine, if there is enough sugar, the yeasts can make so much
alcohol that they kill themselves. This is just one example, but the same
thing can happen with acids and other substances as well.
Both of these examples refer to well-mixed cultures of single cells. In
industry, this is the most common type, and the most dense kind of cell
culturing. It is not the only type of culture, though. Many scientists are
working on making structured groups of cells -- like might be used for an
artificial tissue or organ. In these cases, the rules are completely
different. You have very dense cultures (the cells are all touching one
another), and you have to find away to get nutrients and oxygen to them
without being able to stir them. Scientists are working on artificial
vessels to feed the cells, while others just immerse the clump of cells in a
tank full of medium. This is an ongoing research problem that scientists are
still trying to solve.
Each type of cell and each type of culture is different -- some cells can be
very dense, while others cannot be. There's no single number that applies to
Hope this helps,
Mammalian cells are limited to approximately 50 cell divisions. This number
is known as the Hayflick number, named after the scientist who observed this
phenomenon. This number obviously does not apply to germ (sex) cells,
otherwise we wouldn't be here. This number also does not apply to cancer
cells. HeLa cells (a human cervical cancer cell line) has been growing in
tissue culture since 1950. The currently accepted theory to explain this limit
is that every time a chromosome duplicates, some DNA from the tips of
hromosomes (the telomeres) is lost which eventually prevents the chromosome
Ron Baker, Ph.D.
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Update: June 2012