HepG2 cell line and the common lines beside it: what each models

Using a workhorse line honestly

1. State what the line models. Write down which property of the biology the line is standing in for. If that property is one the line is known to have lost, the experiment needs a different system or an explicit caveat.
2. Authenticate and screen on arrival. Short tandem repeat profiling against a reference and a contamination screen, before the line joins the culture area. The most used lines are also the most misidentified, precisely because they are shared so widely.
3. Fix a passage window. These lines drift with passage, and the drift is faster in some than others. Bank early at low passage, work from the bank, and record passage number with every experiment.
4. Control the differentiation state. Several of these lines behave very differently depending on confluence, medium and whether they have been differentiated. Fix the protocol and describe the state, because the same line can be two systems.
5. Pair with a primary system where the claim needs it. Where a conclusion depends on physiology the line has lost, a primary or stem cell derived system is what supports it. The line is then the screening tool and the primary system the confirmation.

Convenience has a cost

These lines are used because they are robust, cheap and familiar, and every one of those properties comes from adaptations that took them away from the tissue. That is a reasonable trade when the question suits it.

The failure is not using them; it is using them and describing the result as though it came from the tissue.

Provenance travels badly

A line handed between laboratories for years accumulates undocumented passages, medium changes and occasionally an identity swap. The stock in your freezer may differ substantially from the one in the original publication.

Buy from a repository where possible, authenticate on arrival, and record where the stock came from. It costs little and it is the first question a careful reviewer asks.

An mcf7 cell line, and what to check on arrival

This breast carcinoma line is one of the most widely used and most widely misidentified in the literature, so authentication is the first job: short tandem repeat profiling against the reference, a mycoplasma test, and a passage record from the vial you received. It is oestrogen receptor positive and hormone responsive, which is the reason to use it and the reason the serum matters, since phenol red and steroid in serum both affect it. Keep the passage window narrow, because responsiveness drifts with passage.

A human cardiomyocyte cell line, and why there is not really one

There is no immortalised human cardiomyocyte line that behaves like a cardiomyocyte: proliferating lines lose the contractile phenotype, which is why cardiac work runs on cardiomyocytes differentiated from induced pluripotent stem cells, on primary cells with a short useful life, or on animal-derived preparations. Differentiated cells arrive as a defined lot with a purity figure and a maturation state, and both belong in the method, since electrophysiology and drug response depend on how mature the cells are rather than on the donor alone.

hct116 cells and the colorectal panel

hct116 cells are a microsatellite unstable colorectal line, which is exactly why they are paired with a stable line in any experiment about mismatch repair or immune recognition. They grow fast and transfect well, and their near diploid karyotype is unusual among tumour lines, which makes them a common host for gene editing.

An a498 cell line and the renal models

An a498 cell line is a renal carcinoma line carrying the pathway lesion that subtype is defined by, so it is used where a model has to reproduce that biology rather than merely grow. Lines from this tumour type differ in that lesion, which is why the panel rather than one line is what an experiment uses.

An h1975 cell line and the mutation it carries

An h1975 cell line carries the resistance mutation in the receptor that defines a whole class of inhibitor experiments, which is why it is the standard resistant comparator against a sensitive line. A result on it means little without the matched sensitive line on the same plate.

An sw48 cell line and isogenic panels

An sw48 cell line is used as the parental background for isogenic sets in which one mutation is engineered in, and the value of such a set is that everything except the edit is held constant. Isogenic lines have to be authenticated like any other, since a background drifts over passage.

Common questions

Are HepG2 cells a model of liver?

Partly. It retains some hepatic functions and has lost much of the drug metabolising capacity of primary hepatocytes, so it models some questions well and metabolism poorly. Say which you are relying on.

Why authenticate a line everybody uses?

Because widely shared lines are the most likely to have been swapped somewhere in the chain, and morphology will not reveal it. Profiling is inexpensive and settles it permanently for that stock.

Do these lines need a passage limit?

Yes. They drift, and a study spanning a wide passage range has an uncontrolled variable. Bank at low passage and return to the bank rather than extending a culture indefinitely.

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