We’ll appreciate to receive your suggestion about the cell line type to employ in an in vitro experimental model of chronic/acute pancreatitis.
Our starting idea to employ human cell line was hampered by the fact that only primary h-acinar cell are available with a limited life span, which does not fit with the experiment length and reproducibility.
Immortalized h-Ducts cells are available, even if they play a critical role in pancreatitis, aren’t the critical target of the process. In addition , even if in theory they can overexpress some receptors in pancreatitis, the limited number and weight cannot allow to translate in vitro results to a possible radiotracer application in vivo.
Rat-AR42J cell line seems to be the only choice, but it comes from a rat transplantable tumour, is tumorigenic in nude mice and need glucocorticoid stimulus to resemble exocrine function.
There is somebody able to suggest possible alternatives?
As you said, your options are definitely quite limited. The only good acinar cell line is indeed AR42J but in my experience the requirement for GC stimulus is not accurate. You can grow AR42J in F12K supplemented with 20% FBS and they will produce digestive enzymes just fine (amylase, chymotrypsin).
The drawback of of using primary cells is that as soon as primary acinar cells are removed from the context of the pancreas tissue they stop producing protein and can only be used for the first 8-16 hours, after which they are effectively useless.
Mouse primary pancreatic acinar cells are easy enough to produce and can be done on a daily basis for your experiments. This is what I had to do. You can get quite a lot of acinar cells from a single large mouse (12-16 week old CD-1, for example).
The mice have to be at least 12 weeks old or the pancreas will be too immature and will not have a good physiological phenotype for acinar cells.
On last point: AR42J cells won't die upon stimulation with caerulein if you are trying to simulate Cer-AP in vitro, whereas primary acinar cells will (though of course the cell death is quite low in this model - only about 10-15%).
Hello,
I agree with the previous comment. I had been working with the primary mouse acinar cells. It’s an easy and reproducible protocol. The primary acinar cell explants can be planted on a collagen coated well (For a longer period of 7days) and even in suspension culture (For a short period of max.3days). The collagen based culture is most preferable for studying the ADM formation upon external stimulus. If we use suspension culture, we can even measure the secreted inflammatory factors in the medium by ELISA.
Additionally you can have a look at 266-6 cells (in ATCC). These cells mimic the normal acinar cells a lot. They do express amylase in normal condition and upon EGF stimulus they start expressing ductal markers while there is a reduction in amylase expression (detectable in WB). But please do remember that these cells harbor Kras mutation.
I used them in a complete different context. I am not exactly sure if it's applicable for your need.
Jordan hello and thanks for your valuable answer. We are thninking about producing digestive enzymes from the AR42J cells too by using a construct which includes the cds of trypsinogen and amylase.. Do you really think that GC treatment (eg CCK) is not essential? And that if we only use the F12K medium we can obtain the enzymes? Also, we need to know whether trypsinogen or trypsin is obtained from the cells in ZG or in free form..
We also thought about simulating a pancreatitis situation (prolonged exposure to CCK, high Ca2+, acidic pH) in order for the enzymed to be produced at large scale and in their active form. Is that a better scenario?
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