Successful therapy for radiation-induced salivary gland (SG) hypofunction is currently unavailable; however, tissue-specific stem cells are expected to be promising candidates for SG regeneration. Here, we present our method for the establishment of single cell-derived clonal stem cells from mouse SGs and describe their characteristics. Salivary gland-derived clonal stem cells (SGSCs) were isolated and expanded in vitro by a modified subfractionation culture method. The properties of SGSCs were examined with respect to their marker expression, gene expression, differentiation potential, and in vitro immunosuppressive activity relative to bone marrow-derived mesenchymal stem cells (BM-MSCs). SGSCs appeared to largely share the characteristics of BM-MSCs based on their marker expression, whereas they differentially expressed some genes, including AQP5, E-Cadherin, Laminin, ZO-1, and COL4. SGSCs showed the ability to differentiate into fat, bone, and cartilage cell types, as well as into α-amylase-producing and hepatocyte-like cells after appropriate induction. The in vitro immunosuppressive activity of SGSCs was found to be more potent than that of BM-MSCs. These results showed that SGSCs possess the properties of MSCs with some differential gene expression and they are salivary-specific stem cells with both epithelial and mesenchymal properties. The biological functions of SGSCs and their relevance to SG epithelial progenitor cells require further investigation.
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References
1.
JensenS.B., PedersenA.M., VissinkA., AndersenE., BrownC.G., DaviesA.N., DutilhJ., FultonJ.S., JankovicL., LopesN.N., MelloA.L., MunizL.V., Murdoch-KinchC.A., NairR.G., NapenasJ.J., Nogueira-RodriguesA., SaundersD., StirlingB., von BultzingslowenI., WeikelD.S., EltingL.S., SpijkervetF.K., and BrennanM.T.A systematic review of salivary gland hypofunction and xerostomia induced by cancer therapies: prevalence, severity and impact on quality of life. Support Care Cancer, 18, 1039, 2010.
2.
VissinkA., JansmaJ., SpijkervetF.K., BurlageF.R., and CoppesR.P.Oral sequelae of head and neck radiotherapy. Crit Rev Oral Biol Med, 14, 199, 2003.
3.
ChambersM.S., GardenA.S., KiesM.S., and MartinJ.W.Radiation-induced xerostomia in patients with head and neck cancer: pathogenesis, impact on quality of life, and management. Head Neck, 26, 796, 2004.
4.
LombaertI.M., WierengaP.K., KokT., KampingaH.H., deHaanG., and CoppesR.P.Mobilization of bone marrow stem cells by granulocyte colony-stimulating factor ameliorates radiation-induced damage to salivary glands. Clin Cancer Res, 12, 1804, 2006.
5.
LombaertI.M., BrunstingJ.F., WierengaP.K., FaberH., StokmanM.A., KokT., VisserW.H., KampingaH.H., de HaanG., and CoppesR.P.Rescue of salivary gland function after stem cell transplantation in irradiated glands. PLoS One, 3, e2063, 2008.
6.
SumitaY., LiuY., KhaliliS., MariaO.M., XiaD., KeyS., CotrimA.P., MezeyE., and TranS.D.Bone marrow-derived cells rescue salivary gland function in mice with head and neck irradiation. Int J Biochem Cell Biol, 43, 80, 2011.
7.
LimJ.Y., YiT., ChoiJ.S., JangY.H., LeeS., KimH.J., SongS.U., and KimY.M.Intraglandular transplantation of bone marrow-derived clonal mesenchymal stem cells for amelioration of post-irradiation salivary gland damage. Oral Oncol, 49, 136, 2013.
8.
LinC.Y., ChangF.H., ChenC.Y., HuangC.Y., HuF.C., HuangW.K., JuS.S., and ChenM.H.Cell therapy for salivary gland regeneration. J Dent Res, 90, 341, 2011.
9.
KojimaT., KanemaruS., HiranoS., TateyaI., OhnoS., NakamuraT., and ItoJ.Regeneration of radiation damaged salivary glands with adipose-derived stromal cells. Laryngoscope, 121, 1864, 2011.
10.
LimJ.Y., RaJ.C., ShinI.S., JangY.H., AnH.Y., ChoiJ.S., KimW.C., and KimY.M.Systemic transplantation of human adipose tissue-derived mesenchymal stem cells for the regeneration of irradiation-induced salivary gland damage. PLoS One, 8, e71167, 2013.
11.
OkumuraK., NakamuraK., HisatomiY., NaganoK., TanakaY., TeradaK., SugiyamaT., UmeyamaK., MatsumotoK., YamamotoT., and EndoF.Salivary gland progenitor cells induced by duct ligation differentiate into hepatic and pancreatic lineages. Hepatology, 38, 104, 2003.
12.
HisatomiY., OkumuraK., NakamuraK., MatsumotoS., SatohA., NaganoK., YamamotoT., and EndoF.Flow cytometric isolation of endodermal progenitors from mouse salivary gland differentiate into hepatic and pancreatic lineages. Hepatology, 39, 667, 2004.
13.
KishiT., TakaoT., FujitaK., and TaniguchiH.Clonal proliferation of multipotent stem/progenitor cells in the neonatal and adult salivary glands. Biochem Biophys Res Commun, 340, 544, 2006.
14.
SatoA., OkumuraK., MatsumotoS., HattoriK., HattoriS., ShinoharaM., and EndoF.Isolation, tissue localization, and cellular characterization of progenitors derived from adult human salivary glands. Cloning Stem Cells, 9, 191, 2007.
15.
RotterN., OderJ., SchlenkeP., LindnerU., BohrnsenF., KramerJ., RohwedelJ., HussR., BrandauS., WollenbergB., and LangS.Isolation and characterization of adult stem cells from human salivary glands. Stem Cells Dev, 17, 509, 2008.
16.
GorjupE., DannerS., RotterN., HabermannJ., BrassatU., BrummendorfT.H., WienS., MeyerhansA., WollenbergB., KruseC., and von BriesenH.Glandular tissue from human pancreas and salivary gland yields similar stem cell populations. Eur J Cell Biol, 88, 409, 2009.
17.
SongS.U., KimC.S., YoonS.P., KimS.K., LeeM.H., KangJ.S., ChoiG.S., MoonS.H., ChoiM.S., ChoY.K., and SonB.K.Variations of clonal marrow stem cell lines established from human bone marrow in surface epitopes, differentiation potential, gene expression, and cytokine secretion. Stem Cells Dev, 17, 451, 2008.
18.
JeonM.S., YiT.G., LimH.J., MoonS.H., LeeM.H., KangJ.S., KimC.S., LeeD.H., and SongS.U.Characterization of mouse clonal mesenchymal stem cell lines established by subfractionation culturing method. World J Stem Cells, 3, 70, 2011.
19.
Rugel-StahlA., ElliottM.E., and OvittC.E.Ascl3 marks adult progenitor cells of the mouse salivary gland. Stem Cell Res, 8, 379, 2012.
20.
BullardT., KoekL., RoztocilE., KingsleyP.D., MirelsL., and OvittC.E.Ascl3 expression marks a progenitor population of both acinar and ductal cells in mouse salivary glands. Dev Biol, 320, 72, 2008.
21.
FengJ., van der ZwaagM., StokmanM.A., van OsR., and CoppesR.P.Isolation and characterization of human salivary gland cells for stem cell transplantation to reduce radiation-induced hyposalivation. Radiother Oncol, 92, 466, 2009.
22.
DavidR., ShaiE., AframianD.J., and PalmonA.Isolation and cultivation of integrin alpha(6)beta(1)-expressing salivary gland graft cells: a model for use with an artificial salivary gland. Tissue Eng Part A, 14, 331, 2008.
23.
NanduriL.S., MaimetsM., PringleS.A., van der ZwaagM., van OsR.P., and CoppesR.P.Regeneration of irradiated salivary glands with stem cell marker expressing cells. Radiother Oncol, 99, 367, 2011.
24.
LombaertI.M., and HoffmanM.P.Epithelial stem/progenitor cells in the embryonic mouse submandibular gland. Front Oral Biol, 14, 90, 2010.
25.
OkumuraK., ShinoharaM., and EndoF.Capability of tissue stem cells to organize into salivary rudiments. Stem Cells Int, 2012, 502136, 2012.
26.
YanivA., NeumannY., DavidR., Stiubea-CohenR., OrbachY., LangS., RotterN., Dvir-GinzbergM., AframianD.J., and PalmonA.Establishment of immortal multipotent rat salivary progenitor cell line toward salivary gland regeneration. Tissue Eng Part C Methods, 17, 69, 2011.
