The Hayflick Limit​ ​ ​ ​ ​ Name: ______________________________________

Essential Question: Why do cells need to die?

Pre-thinking: Answer the following questions about cell growth in the box below.
What does it mean to die of “natural causes” or to die of “old age?”

Why does this happen?

Is there any living thing that doesn’t die, that can live forever?

​
Part 1: Analyzing research data-Exploring the Hayflick Limit Theory ​
Read the following excerpts about the Hayflick Limit Theory (Also called the Hayflick effect) As you read, box
any words you do not understand which you also think may be important to know in order to get the ‘gist’ of
the articles. Look up the meaning of those words. Also look for the researchers’ claims and the evidence
stated which supports those claims. Put a circle around a claim; underline each piece of evidence. Once you
have the excerpts, answer the following questions.

For the excerpts provided, answer the following questions:

1.​ What claims are made in the data provided?

2.​ What evidence can you list to support those claims? Provide at least three (3) pieces of evidence.

a.​ Evidence:

i.​ Where it was found?

b.​ Evidence:

i.​ Where it was found?

c.​ Evidence:

i.​ Where it was found?

3.​ What conclusion can you assume or draw from the articles and data?​

4.​ Write a question about these findings. What more do you want to know? Or what else might you
consider testing?
Initial experiment in 1961​ ​ ​ ​ ​ ​ ​ ​ ​ Student Data 1

Dr. Leonard Hayflick began to think that the cell division processes had a counting mechanism. Working with
Paul Moorhead, a cytogeneticist, he designed an experiment to test Carrel's theory of cell division (the
leading theory of cell division at the time which stated that human cells were immortal and could divide
indefinitely).
The experiment proceeded as follows. Hayflick and Moorhead mixed equal numbers of normal human male
fibroblasts that had divided many times (cells at the 40th population doubling) with female fibroblasts that
had divided only a few times (cells at the 10th population doubling). Unmixed cell populations were kept as
controls. When the male control culture stopped dividing, the mixed culture was examined and only female
cells were found. This showed that the old male cells remembered they were old, even when surrounded by
young cells, and that technical errors or contaminating viruses were unlikely explanations as to why only
the male cell component had died.[1][4] The cells had stopped dividing and had become senescent based
purely upon how many times the cell had divided.

Follow up experiment 1962​ ​ ​ ​ ​ ​ ​ ​ ​ Student Data 2

​
In 1962 two cell biologists, Dr. Hayflick and Dr. Moorehead, made one of the greatest contributions to the
history of cellular biology by demonstrating the senescence of cultured human cells. Hayflick theorized that
the aging process was controlled by a biological clock contained within each living cell. The 1961 studies
concluded that human fibroblast cells (lung, skin, muscle, heart) have a limited life span. They divided
approximately 50 times over a period of years and then suddenly stopped. Nutrition seemed to have an
effect on the rate of cell division: overfed cells made up to 50 divisions in a year, while underfed cells took
up to three times as long as normal cells to make divisions. Alterations and degenerations occurred within
some cells before they reached their growth limit. The most evident changes took place in the cell
organelles, membranes and genetic material. This improper functioning of cells and loss of cells in organs
and tissues may be responsible for the effects of aging.

Follow up / continuation experiment by Hayflick 1985​ ​ ​ ​ ​ Student Data 3

The normal human fibroblast cell strain WI-38 was established in 1962 from fetal lung, and several hundred
ampules of these cells were frozen in liquid nitrogen at that time. These ampules have been reconstituted
periodically and shown to be capable of replication. This represents the longest period of time that a
normal human cell has ever been frozen. Normal human fetal cell strains such as WI-38 have the capacity
to double only about 50 times. If cultures are frozen at various population doublings, the number of
doublings remaining after reconstitution is equal to 50 minus the number of doublings that occurred prior
to freezing. The memory of the cells has been found to be accurate after 23 years of preservation in liquid
nitrogen. Normal human cells incur many physiologic decrements that herald the approach of their failure
to divide. Many of these functional decrements are identical to decrements found in humans as they age.
Thus it is likely that these decrements are also the precursors of age changes in vivo. The finite replicative
capacity of normal cells is never seen to occur in vivo because aging and death of the individual occurs well
before the doubling limit is reached.
Data Table summarizing other research results​ ​ ​ ​ ​ ​ Student Data 4

Table 1: The average number of cell doublings (or rounds of cell division) in human fetal cells in vitro.

Normal Cells​ Normal Cells in 3% Oxygen​ Cancer cells

In 20% Oxygen (equal to internal environment of humans)

Average number of cell doublings 50 70 No limit

Data Table summarizing other research results​ ​ ​ ​ ​ ​ Student Data 5

​
Table 2: The average number of cell doublings (or rounds of cell division) in MRC-5 cells in vitro

Cryogenic (frozen) Storage Time (months, or years as noted) Average number of cell doublings - ​
(frozen cells started at 7 divisions o month at o divisions)

0 (unfrozen newly derived cells at 0 prior divisions) 48

1 mo. 43

9 mo 42

14 mo 45

18 mo 46

23 mo 44

35 mo 45

23 years 50 minus number of cell divisions prior to cryogenics (freezing)

Part 2: ​ ​ ​ ​ ​ ​ ​ ​ ​ Name: ________________________

Background: Read the following new information.

Cell processes are the result of complex interactions between a cell’s genetic code (DNA) and its environment.
Cells grow, divide, and die in response to cellular and extracellular signals. Sometimes cells are damaged
beyond repair by environmental factors and undergo a programmed cell death - apoptosis. Some cells take
advantage of apoptosis to die for the greater good of the organism.

Before a cell divides, it must make copies of its DNA and all the cellular organelles needed for the daughter
cells. Each time DNA replication occurs, mutations occur. The cell has mechanisms for repairing both mistakes
in DNA replication and other non-replication mutations that occur, but they are not foolproof. Even these
mechanisms for repair are controlled by proteins that are produced from genes that are susceptible to
mutations. If mutations occur in any of the key molecules along the repair pathways, cancer susceptibility
increases as mutation rates accelerate.

In addition, with each cell division the non-coding ends of chromosomes, called telomeres, get shorter and
shorter. People have often compared telomeres to the plastic tips on shoelaces. If they are gone, the shoelace
starts to fray. Every time a cell replicates its DNA, it loses some of the telomeric DNA on both ends. Eventually,
these telomeres are completely gone, and important coding DNA will start to not be copied, leading to cell
death. This, in part, places a limit on the life of a cell. Telomere length alone does not account for the aging
processes of cells or of an organism. Instead, the aging process is a complex interaction of intracellular events
that are still not well understood. Cancer cells, however, have found a way around this limit. They have an
enzyme called telomerase that repairs the length of telomeres, allowing for unlimited cell divisions. Cells that
have telomerase can become immortal.

There are other noncancerous cells, however, that also produce telomerase –stem cells for example – so the
characteristics of cancer cells must go beyond this. There are checkpoints in a cell’s life cycle to make certain
that the cell is on track for DNA replication and for cell division. Mutations in the proteins that regulate these
checkpoints are likely candidates for developing cancer.

In fact, cancer is not as simple as inheriting one mutation in a single gene; it involves multiple mutations in
multiple genes.

Once you have read the selection, answer the following questions:

Why do cells have to die?

What if they don’t die?

What could keep them from dying? Provide evidence for your answer.​