Establishment of functional acinar-like cultures from human salivary glands

J Dent Res. 2015 Feb;94(2):304-11. doi: 10.1177/0022034514559251. Epub 2014 Nov 21.

Abstract

Disorders of human salivary glands resulting from therapeutic radiation treatment for head and neck cancers or from the autoimmune disease Sjögren syndrome (SS) frequently result in the reduction or complete loss of saliva secretion. Such irreversible dysfunction of the salivary glands is due to the impairment of acinar cells, the major glandular cells of protein, salt secretion, and fluid movement. Availability of primary epithelial cells from human salivary gland tissue is critical for studying the underlying mechanisms of these irreversible disorders. We applied 2 culture system techniques on human minor salivary gland epithelial cells (phmSG) and optimized the growth conditions to achieve the maintenance of phmSG in an acinar-like phenotype. These phmSG cells exhibited progenitor cell markers (keratin 5 and nanog) as well as acinar-specific markers-namely, α-amylase, cystatin C, TMEM16A, and NKCC1. Importantly, with an increase of the calcium concentration in the growth medium, these phmSG cells were further promoted to acinar-like cells in vitro, as indicated by an increase in AQP5 expression. In addition, these phmSG cells also demonstrated functional calcium mobilization, formation of epithelial monolayer with high transepithelial electrical resistance (TER), and polarized secretion of α-amylase secretion after β-adrenergic receptor stimulation. Taken together, suitable growth conditions have been established to isolate and support culture of acinar-like cells from the human salivary gland. These primary epithelial cells can be useful for study of molecular mechanisms involved in regulating the function of acinar cells and in the loss of salivary gland function in patients.

Keywords: Sjögren’s syndrome; acinar cells; calcium mobilization; explant culture; primary cell culture; primary epithelial cells.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Anoctamin-1
  • Aquaporin 5 / analysis
  • Calcium / pharmacology
  • Calcium Signaling / physiology
  • Cell Adhesion Molecules / analysis
  • Cell Culture Techniques
  • Cell Differentiation / drug effects
  • Chloride Channels / analysis
  • Culture Media
  • Cystatin C / analysis
  • Electric Impedance
  • Epithelial Cells / cytology
  • Homeodomain Proteins / analysis
  • Humans
  • Keratin-5 / analysis
  • Membrane Proteins / analysis
  • Nanog Homeobox Protein
  • Neoplasm Proteins / analysis
  • Phenotype
  • Receptors, Adrenergic, beta / drug effects
  • Salivary Glands, Minor / cytology*
  • Solute Carrier Family 12, Member 2 / analysis
  • Stem Cells / cytology
  • Stromal Interaction Molecule 1
  • Stromal Interaction Molecule 2
  • TRPC Cation Channels / analysis
  • Tight Junctions / ultrastructure
  • alpha-Amylases / analysis

Substances

  • ANO1 protein, human
  • AQP5 protein, human
  • Anoctamin-1
  • Aquaporin 5
  • CST3 protein, human
  • Cell Adhesion Molecules
  • Chloride Channels
  • Culture Media
  • Cystatin C
  • Homeodomain Proteins
  • KRT5 protein, human
  • Keratin-5
  • Membrane Proteins
  • NANOG protein, human
  • Nanog Homeobox Protein
  • Neoplasm Proteins
  • Receptors, Adrenergic, beta
  • SLC12A2 protein, human
  • STIM1 protein, human
  • STIM2 protein, human
  • Solute Carrier Family 12, Member 2
  • Stromal Interaction Molecule 1
  • Stromal Interaction Molecule 2
  • TRPC Cation Channels
  • transient receptor potential cation channel, subfamily C, member 1
  • alpha-Amylases
  • Calcium