title: Cryopreserved mouse pancreatic acinar cells from long-term explant outgrowth cultures maintain their secretory phenotype after thawing.
abstract: OBJECTIVE
  We recently reported an explant outgrowth culture method for obtaining functionally competent mouse pancreatic acinar cells for long-term in vitro purposes. The aim of the present study was to explore the possibility of cryostoring these cells without loss of functional differentiation.
  
  
  METHODS
  Acinar cells prepared by the explant outgrowth method were cryopreserved using a DMSO-based protocol and stored in liquid nitrogen for 4 weeks. The following characteristics were compared in cryopreserved and parallel non-frozen cell preparations: cell viability and recovery, amylase content in viable cells before culture, basal and stimulated amylase release in culture and the ability of the cells to form glandular structures in Matrigel.
  
  
  RESULTS
  Immediate post-thaw viability of the cells was similar to that of freshly isolated cells. Approximately 53% of viable cells frozen were recovered after thawing. Intracellular amylase content was identical in frozen and non-frozen cells. Cryopreserved cells maintained their ability to secrete amylase and to respond to caerulein stimulation in 4-day secondary cultures. They also were observed to form amylase-expressing glandular structures in three-dimensional cultures in Matrigel in a similar manner as non-frozen cells.
  
  
  CONCLUSIONS
  This study shows that pancreatic acinar cells can be cryopreserved for long-term storage in liquid nitrogen without dedifferentiation. Successful cryopreservation helps to refine the experimental use of primary acinar cells by enabling their banking for on-demand utilization.
authors.affiliation: Department of Gastroenterology and Alimentary Tract Surgery and Tampere Pancreas Laboratory, Tampere University Hospital, Teiskontie 35, FIN-33521 Tampere, Finland.
authors.forename: Merja
authors.initials: M
authors.lastname: Bläuer
authors.forename: Juhani
authors.initials: J
authors.lastname: Sand
authors.forename: Johanna
authors.initials: J
authors.lastname: Laukkarinen
mesh_terms: D061354:Acinar Cells / Q000378:metabolism* / Q000502:physiology*; D000681:Amylases / Q000032:analysis; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005615:Freezing; D005990:Glycerol / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009924:Organ Culture Techniques; D010179:Pancreas / Q000166:cytology* / Q000378:metabolism*; D010641:Phenotype; D061251:Primary Cell Culture
chemical_list: D003451:Cryoprotective Agents; D007770:L-Lactate Dehydrogenase; D000681:Amylases; D005990:Glycerol; D004121:Dimethyl Sulfoxide
keywords: Cryostorage; DMSO; Exocrine pancreas; In vitro; Long-term culture