title: Long-term cryopreservation model of rat salivary gland stem cells for future therapy in irradiated head and neck cancer patients.
abstract: Irradiated head and neck cancer patients suffer from irreversible loss of salivary gland (SG) function, along with significant morbidity and compromised quality of life. To date there is no biologically-based treatment for this distress. Adult salivary gland stem cells are promising candidates for autologous transplantation therapy in the context of tissue-engineered artificial SGs or direct cell therapy. The major restrictions in handling such cells are their limited lifespan during in vitro cultivation, resulting in a narrow time-window for implantation and a risk of tumorigenic changes during culture. To overcome these difficulties, we tested in a rat model the possibility of establishing a personal/autologous SG stem cell bank. SG's integrin-α6β1-expressing cells were shown to hold a subpopulation of SG-specific progenitor-cells. Explanted and cultured single cell-originated clones were cryopreserved for up to 3 years and shown to exhibit genetic and functional stability similar to noncryopreserved cells, as was emphasized by soft agar assay, division potential assessment, flow cytometric analysis, real-time reverse transcriptase-polymerase chain reaction, in vitro three-dimensional differentiation assay, and immunofluorescence confocal microscopy. Future integration of the novel strategies presented herein to a clinical therapeutic model will allow safe preservation until transplantation and repeated transplantation if needed. These tools open a new venue for adult autologous stem-cell transplantation-based SG regeneration.
authors.affiliation: Faculty of Dental Medicine, Institute of Dental Sciences, The Hebrew University, Jerusalem, Israel .
authors.forename: Yoav
authors.initials: Y
authors.lastname: Neumann
authors.forename: Ran
authors.initials: R
authors.lastname: David
authors.forename: Raluca
authors.initials: R
authors.lastname: Stiubea-Cohen
authors.forename: Yoav
authors.initials: Y
authors.lastname: Orbach
authors.forename: Doron J
authors.initials: DJ
authors.lastname: Aframian
authors.forename: Aaron
authors.initials: A
authors.lastname: Palmon
mesh_terms: D016475:3T3 Cells; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D017690:Cell Transplantation / Q000379:methods*; D015925:Cryopreservation; D006258:Head and Neck Neoplasms / Q000532:radiotherapy* / Q000628:therapy; D006801:Humans; D039121:Integrin alpha6beta1 / Q000096:biosynthesis; D008297:Male; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D051381:Rats; D012038:Regeneration; D012469:Salivary Glands / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods
chemical_list: D039121:Integrin alpha6beta1