Re-engineering primary epithelial cells from rhesus monkey parotid glands for use in developing an artificial salivary gland.
There is no satisfactory conventional treatment for patients who experience irreversible salivary gland damage after therapeutic radiation for head and neck cancer or because of Sjögren's syndrome. Additionally, if most parenchyma is lost, these patients also are not candidates for evolving gene transfer strategies. To help such patients, several years ago we began to develop an artificial salivary gland. In the present study, we used a non-human primate tissue source, parotid glands from rhesus monkeys, to obtain potential autologous graft cells for development of a prototype device for in situ testing. Herein, we present 3 major findings. First, we show that primary cultures of rhesus parotid gland (RPG) cells are capable of attaining a polarized orientation, with Na(+)/K(+)-adenosine triphosphatase, zonula occludens-1, and claudin-1 distributed in specific domains appropriate for epithelial cells. Second, we show that RPG cells exhibit 2 essential epithelial functions required for graft cells in an artificial salivary gland device (i.e., an effective barrier to paracellular water flow and the generation of a moderate transepithelial electrical resistance). Third, we show that RPG cells can express functional water channels, capable of mediating directional fluid movement, after transduction by adenoviral and adeno-associated virus type 2 vectors. Together these results demonstrate that it is feasible to individually prepare RPG cells for eventual use in a prototype artificial salivary gland.
Tran
Simon D
SD
Gene Therapy and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland 20892-1190, USA.
Sugito
Takayuki
T
Dipasquale
Giovanni
G
Cotrim
Ana P
AP
Bandyopadhyay
Bidhan C
BC
Riddle
Kathryn
K
Mooney
David
D
Kok
Marc R
MR
Chiorini
John A
JA
Baum
Bruce J
BJ
D000818:Animals; D001705:Bioprosthesis; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology* / Q000502:physiology; D004867:Equipment Design; D019544:Equipment Failure Analysis; D008253:Macaca mulatta; D009061:Mouth Mucosa / Q000166:cytology / Q000502:physiology; D009924:Organ Culture Techniques / Q000379:methods*; D010306:Parotid Gland / Q000166:cytology* / Q000502:physiology; D012469:Salivary Glands / Q000166:cytology* / Q000254:growth & development*; D023822:Tissue Engineering / Q000379:methods*