Primary Acinar Cell Isolation: A Method to Isolate Acinar Cells from Mouse Pancreas

Abstract

Source: Fleming Martinez, A. K., et al. Mimicking and Manipulating Pancreatic Acinar-to-Ductal Metaplasia in 3-dimensional Cell Culture. J. Vis. Exp. (2019).

This video describes the technique used for isolating pancreatic acinar cells--exocrine cells--containing a variety of digestive enzymes. Primary acinar cells are useful in the ex vivo study of acinar-to-ductal metaplasia (ADM)--an early event in developing pancreatic cancer.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the jove veterinary review board.

1. Acinar Cell Isolation

1. Sacrifice the mouse via CO₂ induction, and perform cervical dislocation. Immediately dissect the pancreas. To dissect the pancreas, first pin the paws of the mouse to the polystyrene lid, orient the mouse such that the tail is facing the researcher, and spray the abdomen with 70% ethanol.
   NOTE: The mouse used in the representative results was a 12-week old non-transgeni....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
37 °C shaking incubator	Thermo Scientific	SHKE4000-7
5% CO2, 37 °C Incubator	NUAIRE	NU-5500
50 ml tubes	Falcon	352070
Absorbent pad, 20" x 24"	Fisherbrand	1420662
BD Precision Glide Needle 21G x 1/2	Fisher Scientific	305167	Use as pins for dissection
Beaker, 600 mL	Fisherbrand	FB-101-600
Centrifuge	Beckman Coulter	Allegra X-15R Centrifuge
Collagenase Type I, Clostridium histolyticum	Sigma	C0130-1G	Create a 100 mg/ml solution by dissolving powder in sterile molecular biology grade water. When thawing one aliquot, dilute to 10 mg/ml, filter sterilize and place 1 ml aliquots at -20 °C.
Dexamethasone	Sigma	D1756	Create a 4 mg/ml solution by dissolving powder in methanol, aliquoting and storing at -20 °C.
Ethanol, 200 proof	Decon Laboratories	2701
Fetal Bovine Serum	Sigma	F0926-100mL
Forceps	Fine Science Tools	11002-12
Forceps	Fine Science Tools	91127-12
Hank's Balanced Salt Solution (HBSS), No calcium, No magnesium, No phenol red	Fisher Scientific	SH3048801
Ice bucket, rectangular	Fisher Scientific	07-210-103
LAB GUARD specimen bags (for mouse after dissection)	Minigrip	SBL2X69S
Parafilm	Bemis	PM992	Referred to as 'plastic paraffin film' in the manuscript
PBS	Fisher Scientific	SH30028.02
Penicillin-Streptomycin	ThermoFisher Scientific	15140122
Pipet tips, 10 µl	USA Scientific	1110-3700
Pipet tips, 1000 µl	Olympus Plastics	24-165RL
Pipet tips, 200 µl	USA Scientific	1111-1700
Pipet-Aid	Drummond
PIPETMAN Classic P10, 1-10 μl	Gilson	F144802
PIPETMAN Classic P1000, 200-1000 μl	Gilson	F123602
PIPETMAN Classic P20, 2-20 μl	Gilson	F123600
PIPETMAN Classic P200, 20-200 μl	Gilson	F123601
Pipettes, 10 ml	Falcon	357551
Pipettes, 25 ml	Falcon	357525
Pipettes, 5 ml	Falcon	357543
Polypropylene Mesh, 105 µm	Spectrum Labs	146436
Polypropylene Mesh, 500 µm	Spectrum Labs	146418
Scissors	Fine Science Tools	14568-12
Scissors	Fine Science Tools	14568-13
Sodium Bicarbonate (Fine White Powder)	Fisher Scientific	BP328-500
Sodium Hydroxide	Fisher Scientific	S318-500
Soybean Trypsin Inhibitor	Gibco	17075029
Spatula	Fisherbrand	21-401-10
Steriflip 50 ml, 0.22 micron filters	Millipore	SCGP00525
Type I Rat Tail Collagen	Corning	354236
Waymouth MB 752/1 Medium (powder)	Sigma	W1625-10X1L
Weigh boat, hexagonal, medium	Fisherbrand	02-202-101