Reciprocal stimulation of pancreatic acinar and stellate cells in a novel long-term in vitro co-culture model

• Published in: Pancreatology : official journal of the International Association of Pancreatology (IAP) ... [et al.] 2016-07, Vol.16 (4), p.570-577
• Main Authors: Bläuer, Merja, Laaninen, Matias, Sand, Juhani, Laukkarinen, Johanna
• Format: Article
• Language: English
• Subjects: 3T3 Cells; Acinar Cells - drug effects; Amylases - metabolism; Animals; Apoptosis; Cell Communication; Cell culture; Ceruletide - pharmacology; Co-culture; Coculture Techniques; Cytokines; Dual-chamber model; Endocrinology & Metabolism; Experiments; Fibroblasts; Gastroenterology and Hepatology; Growth factors; HMGB1 Protein - genetics; Homeostasis; Humans; Humoral interactions; In vitro; Male; Mice; Mice, Inbred C57BL; NF-kappa B - analysis; NF-kappa B - metabolism; Pancreatic cancer; Pancreatic cells; Pancreatic Stellate Cells - drug effects; Phenotype; Pro-fibrogenic; Protein expression; Rodents; Short term; Signal transduction; Stimulation, Chemical; Tissue Culture Techniques
• ISSN: 1424-3903; 1424-3911
• DOI: 10.1016/j.pan.2016.03.012

Abstract Background/objectives Pancreatic stellate cells (PSCs) are the key fibrogenic cells in the pancreas. Acinar cell injury is known to trigger PSC activation. To facilitate the experimental analysis of the crosstalk between acinar cells and PSCs, an in vitro system for their long-term co-cultivation was developed. Materials and methods PSCs and acinar cells capable of retaining their secretory phenotype in long-term in vitro culture were obtained from mouse pancreata. A dual-chamber co-culture model was built in 24-well format with acinar cells seeded in the wells and PSCs in tissue culture inserts. Acinar cell-3T3 fibroblast co-cultures served as controls. After 4-day maintenance, the acinar compartment was analyzed for cell morphology, secretory capability, necrosis (HMGB1), apoptosis (TUNEL) and inflammation (NFκB). PSCs were analyzed for migratory activity and extracellular matrix (ECM) protein expression. The results were compared to parallel monocultures. Results Acinar cells in monoculture and in co-culture with fibroblasts exhibited a healthy monolayer arrangement and an ability to respond to 0.1 nM caerulein stimulus by increased amylase release. Co-culture with PSCs caused marked changes in acinar cell morphology and rendered them insensitive to secretagogue stimulus. Activation of NFκB and necrotic changes, but not apoptosis, were identified in co-cultured acinar cells. Co-culture increased the migratory activity and ECM protein expression of PSCs. Conclusions Humoral interactions between acinar and PSCs in co-culture were shown to reciprocally affect their cellular functions. With its two separable cell compartments the co-culture system provides a versatile culture setting that allows independent manipulation and analysis of both cell types.