While mesenchymal stem cells (MSCs) are promising candidates for inner ear hair cell regeneration, to date, there have been no convincing reports indicating whether MSCs can survive in the cochlea for more than a few weeks, as the high levels of potassium (K+) in the endolymph (EL) are thought to be toxic to transplanted stem cells. For conditioning the EL for MSC transplantation, we conducted this in vitro study to examine the effects of artificial EL with altered K+ concentration levels, in the range of 5–153.8 mM, on proliferation, apoptosis, and morphological changes in MSCs derived from various human tissues. Our findings demonstrate that altering the K+ concentration in artificial EL could significantly influence the survival of MSCs in vitro. We discovered that K+ concentrations of 55–130 mM in artificial EL could enhance the survival of MSCs in vitro. However, MSCs exhibited reduced proliferation regardless of K+ concentration.
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References
1.
TucciD, MersonMH and WilsonBS. (2010). A summary of the literature on global hearing impairment: current status and priorities for action. Otol Neurotol, 31:31–41.
2.
WarcholME. (2011). Sensory regeneration in the vertebrate inner ear: differences at the levels of cells and species. Hear Res, 273:72–79.
3.
BrignullHR, RaibleDW and StoneJS. (2009). Feathers and fins: non-mammalian models for hair cell regeneration. Brain Res, 1277:12–23.
4.
StoneJS and CotancheDA. (2007). Hair cell regeneration in the avian auditory epithelium. Int J Dev Biol, 51:633–647.
5.
LiH, CorralesCE, EdgeA and HellerS. (2004). Stem cells as therapy for hearing loss. Trends Mol Med, 10:309–315.
6.
AjalloueyanM, KouhiA, AsgariA, SalemM and HasanalifardM. (2013). Stem cell transplantation in noise induced hearing loss. Int J Pediatr Otorhinolaryngol, 77:469–472.
7.
MullerU and Barr-GillespiePG. (2015). New treatment options for hearing loss. Nat Rev Drug Discov, 14:346–365.
8.
XuYP, ShanXD, LiuYY, PuY, WangCY, TaoQL, DengY, ChengY and FanJP. (2016). Olfactory epithelium neural stem cell implantation restores noise-induced hearing loss in rats. Neurosci Lett, 616:19–25.
9.
KakigiA and TakedaT. (1998). Effect of artificial endolymph injection into the cochlear duct on the endocochlear potential. Hear Res, 116:113–118.
10.
MarcusDC, RokugoM, GeXX and ThalmannR. (1983). Response of cochlear potentials to presumed alterations of ionic conductance: endolymphatic perfusion of barium, valinomycin and nystatin. Hear Res, 12:17–30.
11.
BoddySL, ChenW, Romero-GuevaraR, KottamL, BellantuonoI and RivoltaMN. (2012). Inner ear progenitor cells can be generated in vitro from human bone marrow mesenchymal stem cells. Regen Med, 7:757–767.
12.
KilK, ChoiMY, KongJS, KimWJ and ParkKH. (2016). Regenerative efficacy of mesenchymal stromal cells from human placenta in sensorineural hearing loss. Int J Pediatr Otorhinolaryngol, 91:72–81.
13.
KilK, ChoiMY and ParkKH. (2016). In vitro differentiation of human Wharton's jelly-derived mesenchymal stem cells into auditory hair cells and neurons. J Int Adv Otol, 12:37–42.
14.
JongkamonwiwatN, ZineA and RivoltaMN. (2010). Stem cell based therapy in the inner ear: appropriate donor cell types and routes for transplantation. Curr Drug Targets, 11:888–897.
15.
ParkYH. (2015). Stem cell therapy for sensorineural hearing loss, still alive?. J Audiol Otol, 19:63–67.
16.
ParkYH, WilsonKF, UedaY, Tung WongH, BeyerLA, SwiderskiDL, DolanDF and RaphaelY. (2014). Conditioning the cochlea to facilitate survival and integration of exogenous cells into the auditory epithelium. Mol Ther, 22:873–880.
17.
HildebrandMS, DahlHH, HardmanJ, ColemanB, ShepherdRK and de SilvaMG. (2005). Survival of partially differentiated mouse embryonic stem cells in the scala media of the guinea pig cochlea. J Assoc Res Otolaryngol, 6:341–354.
18.
LangH, SchulteBA, GoddardJC, HedrickM, SchulteJB, WeiL and SchmiedtRA. (2008). Transplantation of mouse embryonic stem cells into the cochlea of an auditory-neuropathy animal model: effects of timing after injury. J Assoc Res Otolaryngol, 9:225–240.
19.
ShiM, LiJ, LiaoL, ChenB, LiB, ChenL, JiaH and ZhaoRC. (2007). Regulation of CXCR4 expression in human mesenchymal stem cells by cytokine treatment: role in homing efficiency in NOD/SCID mice. Haematologica, 92:897–904.
20.
ZhouH, GuoM, BianC, SunZ, YangZ, ZengY, AiH and ZhaoRC. (2010). Efficacy of bone marrow-derived mesenchymal stem cells in the treatment of sclerodermatous chronic graft-versus-host disease: clinical report. Biol Blood Marrow Transplant, 16:403–412.
21.
MakimotoK, TakedaT and SilversteinH. (1980). Species differences in inner ear fluids. Arch Otorhinolaryngol, 228:187–193.
22.
WangemannP and SchachtJ. (1996). Homeostatic mechanisms in the cochlea. In: The Cochlea. DallosP, PopperAN, FayRR, eds. Springer, New York, pp 130–185.
23.
DominiciM, Le BlancK, MuellerI, Slaper-CortenbachI, MariniF, KrauseD, DeansR, KeatingA, ProckopD and HorwitzE. (2006). Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy, 8:315–317.
24.
BakshD, SongL and TuanRS. (2004). Adult mesenchymal stem cells: characterization, differentiation, and application in cell and gene therapy. J Cell Mol Med, 8:301–316.
25.
SakagamiM, HaradaT, SanoM, SakaiS and MatsunagaT. (1987). Quantitative evaluation of pinocytosis of capillaries of the stria vascularis under normal and experimental conditions. Acta Otolaryngol, 103:189–197.
26.
TakeuchiS, AndoM, SatoT and KakigiA. (2001). Three-dimensional and ultrastructural relationships between intermediate cells and capillaries in the gerbil stria vascularis. Hear Res, 155:103–112.
27.
JuhnSK. (1988). Barrier systems in the inner ear. Acta Otolaryngol Suppl, 458:79–83.
28.
ShiX. (2009). Cochlear pericyte responses to acoustic trauma and the involvement of hypoxia-inducible factor-1alpha and vascular endothelial growth factor. Am J Pathol, 174:1692–1704.
29.
ShiX. (2011). Physiopathology of the cochlear microcirculation. Hear Res, 282:10–24.
30.
GaoM, YangHY, LiuTY, GuXJ, HuYE, XuG and TangWC. (2014). [Effects of different acupuncture and moxibustion methods on ultrastructure of gastrocnemius in rats]. Zhongguo Zhen Jiu, 34:261–265.
31.
SwiderskiZ, MiquelJ, MarigoAM and GibsonDI. (2012). Ultrastructure of vitellogenesis and vitellocytes in the trypanorhynch cestode Aporhynchus menezesi, a parasite of the velvet belly lanternshark Etmopterus spinax. C R Biol, 335:573–584.
32.
DallaportaB, HirschT, SusinSA, ZamzamiN, LarochetteN, BrennerC, MarzoI and KroemerG. (1998). Potassium leakage during the apoptotic degradation phase. J Immunol, 160:5605–5615.
33.
PerezGI, MaraveiDV, TrbovichAM, CidlowskiJA, TillyJL and HughesFMJr. (2000). Identification of potassium-dependent and -independent components of the apoptotic machinery in mouse ovarian germ cells and granulosa cells. Biol Reprod, 63:1358–1369.
MazurekB, AmarjargalN, HauptH and GrossJ. (2006). High potassium concentrations protect inner and outer hair cells in the newborn rat culture from ischemia-induced damage. Hear Res, 215:31–38.
36.
LeeMY, HackelbergS, GreenKL, LunghamerKG, KuriokaT, LoomisBR, SwiderskiDL, DuncanRK and RaphaelY. (2017). Survival of human embryonic stem cells implanted in the guinea pig auditory epithelium. Sci Rep, 7:46058.
37.
JiangS, ChowSC, NicoteraP and OrreniusS. (1994). Intracellular Ca2+ signals activate apoptosis in thymocytes: studies using the Ca(2+)-ATPase inhibitor thapsigargin. Exp Cell Res, 212:84–92.
38.
WertzIE and DixitVM. (2000). Characterization of calcium release-activated apoptosis of LNCaP prostate cancer cells. J Biol Chem, 275:11470–11477.
39.
BoyerC, LehouelleurJ and SansA. (1998). Potassium depolarization of mammalian vestibular sensory cells increases [Ca2+]i through voltage-sensitive calcium channels. Eur J Neurosci, 10:971–975.
40.
BoyerC, ArtJJ, DechesneCJ, LehouelleurJ, VautrinJ and SansA. (2001). Contribution of the plasmalemma to Ca2+ homeostasis in hair cells. J Neurosci, 21:2640–2650.
41.
PottorfWJ and ThayerSA. (2002). Transient rise in intracellular calcium produces a long-lasting increase in plasma membrane calcium pump activity in rat sensory neurons. J Neurochem, 83:1002–1008.
42.
OsbornKD, ZaidiA, MandalA, UrbauerRJ and JohnsonCK. (2004). Single-molecule dynamics of the calcium-dependent activation of plasma-membrane Ca2+-ATPase by calmodulin. Biophys J, 87:1892–1899.
43.
PerregauxD and GabelCA. (1994). Interleukin-1 beta maturation and release in response to ATP and nigericin. Evidence that potassium depletion mediated by these agents is a necessary and common feature of their activity. J Biol Chem, 269:15195–15203.
44.
WalevI, ReskeK, PalmerM, ValevaA and BhakdiS. (1995). Potassium-inhibited processing of IL-1 beta in human monocytes. EMBO J, 14:1607–1614.
45.
JacobsonMD and EvanGI. (1994). Apoptosis. Breaking the ICE. Curr Biol, 4:337–340.
46.
JentschTJ. (2000). Neuronal KCNQ potassium channels: physiology and role in disease. Nat Rev Neurosci, 1:21–30.
47.
SchmidtRS. (1963). Independence of the endovestibular potential in homeotherms. J Gen Physiol, 47:371–378.
48.
WangemannP. (2002). K(+) cycling and its regulation in the cochlea and the vestibular labyrinth. Audiol Neurootol, 7:199–205.
49.
WangemannP. (1995). Comparison of ion transport mechanisms between vestibular dark cells and strial marginal cells. Hear Res, 90:149–157.
50.
OffnerFF, DallosP and CheathamMA. (1987). Positive endocochlear potential: mechanism of production by marginal cells of stria vascularis. Hear Res, 29:117–124.
51.
TakeuchiS, AndoM and KakigiA. (2000). Mechanism generating endocochlear potential: role played by intermediate cells in stria vascularis. Biophys J, 79:2572–2582.
52.
AndoM and TakeuchiS. (1999). Immunological identification of an inward rectifier K+ channel (Kir4.1) in the intermediate cell (melanocyte) of the cochlear stria vascularis of gerbils and rats. Cell Tissue Res, 298:179–183.
53.
YanglinP, LinaZ, ZhiguoL, NaL, HaifengJ, GuoyunZ, JieL, JunW, TaoL, et al. (2007). KCNE2, a down-regulated gene identified by in silico analysis, suppressed proliferation of gastric cancer cells. Cancer Lett, 246:129–138.
54.
YaoX and KwanHY. (1999). Activity of voltage-gated K+ channels is associated with cell proliferation and Ca2+ influx in carcinoma cells of colon cancer. Life Sci, 65:55–62.
55.
BeetonC, PenningtonMW, WulffH, SinghS, NugentD, CrossleyG, KhaytinI, CalabresiPA, ChenCY, GutmanGA and ChandyKG. (2005). Targeting effector memory T cells with a selective peptide inhibitor of Kv1.3 channels for therapy of autoimmune diseases. Mol Pharmacol, 67:1369–1381.
56.
BeetonC, WulffH, BarbariaJ, Clot-FaybesseO, PenningtonM, BernardD, CahalanMD, ChandyKG and BeraudE. (2001). Selective blockade of T lymphocyte K(+) channels ameliorates experimental autoimmune encephalomyelitis, a model for multiple sclerosis. Proc Natl Acad Sci U S A, 98:13942–13947.
57.
JagerH, DrekerT, BuckA, GiehlK, GressT and GrissmerS. (2004). Blockage of intermediate-conductance Ca2+-activated K+ channels inhibit human pancreatic cancer cell growth in vitro. Mol Pharmacol, 65:630–638.
58.
WangSP, WangRH, LuoWY, CuiWY and WangH. (2008). Potassium channel currents in rat mesenchymal stem cells and their possible roles in cell proliferation. Clin Exp Pharmacol Physiol, 35:1077–1084.
59.
McLeanWJ, YinX, LuL, LenzDR, McLeanD, LangerR, KarpJM and EdgeA. (2017). Clonal expansion of Lgr5-positive cells from mammalian cochlea and high-purity generation of sensory hair cells. Cell Rep, 18:1917–1929.
60.
XuJ, SongD, XueZ, GuL, HertzL and PengL. (2013). Requirement of glycogenolysis for uptake of increased extracellular K+ in astrocytes: potential implications for K+ homeostasis and glycogen usage in brain. Neurochem Res, 38:472–485.
61.
DiNuzzoM, MangiaS, MaravigliaB and GioveF. (2013). Regulatory mechanisms for glycogenolysis and K+ uptake in brain astrocytes. Neurochem Int, 63:458–464.
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