Adult stem cells from the dermis would be an attractive cell source for therapeutic purposes as well as studying the process of skin aging. Several studies have reported that human dermal stem/progenitor cells (hDSPCs) with multipotent properties exist within the dermis of adult human skin. However, these cells have not been well characterized, because methods for their isolation or enrichment have not yet been optimized. In the present study, we enriched high side scatter (SSChigh)-hDSPCs from normal human dermal fibroblasts using a structural characteristic, intracellular granularity, as a sorting parameter. The SSChigh-hDSPCs had high in vitro proliferation properties and expressed high levels of SOX2 and S100B, similar to previously identified mouse SOX2+ hair follicle dermal stem cells. The SSChigh-hDSPCs could differentiate into not only mesodermal cell types, for example, adipocytes, chondrocytes, and osteoblasts, but also neuroectodermal cell types, such as neural cells. In addition, the SSChigh-hDSPCs exhibited no significant differences in the expression of nestin, vimentin, SNAI2, TWIST1, versican, and CORIN compared with non-hDSPCs. These cells are therefore different from the previously identified multipotent fibroblasts and skin-derived progenitors. In this study, we suggest that hDSPCs can be enriched by using characteristic of their high intracellular granularity, and these SSChigh-hDSPCs exhibit high in vitro proliferation and differentiation potentials.
Get full access to this article
View all access options for this article.
References
1.
YoungHE, DuplaaC, KatzR, ThompsonT, HawkinsKC, BoevAN, HensonNL, HeatonM, SoodRet al.2005. Adult-derived stem cells and their potential for use in tissue repair and molecular medicine. J Cell Mol Med, 9:753–769.
2.
PereiraRF, HalfordKW, O'HaraMD, LeeperDB, SokolovBP, PollardMD, BagasraO, ProckopDJ. 1995. Cultured adherent cells from marrow can serve as long-lasting precursor cells for bone, cartilage, and lung in irradiated mice. Proc Natl Acad Sci U S A, 92:4857–4861.
3.
PittengerMF, MackayAM, BeckSC, JaiswalRK, DouglasR, MoscaJD, MoormanMA, SimonettiDW, CraigS, MarshakDR. 1999. Multilineage potential of human mesenchymal stem cells. Science, 284:143–147.
JiangY, JahagirdarBN, ReinhardtRL, SchwartzRE, KeeneCD, Ortiz-GonzalezXR, ReyesM, LenvikTet al.2002. Pluripotency of mesenchymal stem cells derived from adult marrow. Nature, 418:41–49.
6.
JiangY, VaessenB, LenvikT, BlackstadM, ReyesM, VerfaillieCM. 2002. Multipotent progenitor cells can be isolated from postnatal murine bone marrow, muscle, and brain. Exp Hematol, 30:896–904.
7.
ZukPA, ZhuM, MizunoH, HuagJ, FutrellJW, KatzAJ, BenhaimP, LorenzHP, HedrickMH. 2001. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng, 7:211–218.
8.
ZukPA, ZhuM, AshjianP, De UgarteDA, HuangJI, MizunoH, AlfonsoZC, FraserJK, BenhaimP, HedrickMH. 2002. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell, 13:4279–4295.
9.
LeeMW, YangMS, ParkJS, KimHC, KimYJ, ChoiJ. 2005. Isolation of mesenchymal stem cells from cryopreserved human umbilical cord blood. Int J Hematol, 81:126–130.
10.
FukuchiY, NakajimaH, SugiyamaD, HiroseI, KitamuraT, TsujiK. 2004. Human placenta-derived cells have mesenchymal stem/progenitor cell potential. Stem Cells, 22:649–658.
11.
BlanpainC, FuchsE. 2006. Epidermal stem cells of the skin. Annu Rev Cell Dev Biol, 22:339–373.
12.
FuchsE. 2009. Finding one's niche in the skin. Cell Stem Cell, 4:499–502.
13.
TomaJG, AkhavanM, FernandesKJ, Barnabé-HeiderF, SadikotA, KaplanDR, MillerFD. 2001. Isolation of multipotent adult stem cells from the dermis of mammalian skin. Nat Cell Biol, 3:778–784.
TomaJG, McKenzieIA, BagliD, MillerFD. 2005. Isolation and characterization of multipotent skin-derived precursors from human skin. Stem Cells, 23:727–737.
16.
ChenFG, ZhangWJ, BiD, LiuW, WeiX, ChenFF, ZhuL, CuiL, CaoY. 2007. Clonal analysis of nestin(−) vimentin(+) multipotent fibroblasts isolated from human dermis. J Cell Sci, 120:2875–2883.
17.
DengY, HuJC, AthanasiouKA. 2007. Isolation and chondroinduction of a dermis isolated, aggrecan-sensitive subpopulation with high chondrogenic potential. Arthritis Rheum, 56:168–176.
18.
BiernaskieJ, ParisM, MorozovaO, FaganBM, MarraM, PevnyL, MillerFD. 2009. SKPs derive from hair follicle precursors and exhibit properties of adult dermal stem cells. Cell Stem Cell, 5:610–623.
19.
DriskellRR, GiangrecoA, JensenKB, MulderKW, WattFM. 2009. Sox2-positive dermal papilla cells specify hair follicle type in mammalian epidermis. Development, 7136:2815–2823.
20.
JunkerJP, SommarP, SkogM, JohnsonH, KratzG. 2010. Adipogenic, chondrogenic and osteogenic differentiation of clonally derived human dermal fibroblasts. Cells Tissues Organs, 191:105–118.
21.
AmohY, LiL, CampilloR, KawaharaK, KatsuokaK, PenmanS, HoffmanRM. 2005. Implanted hair follicle stem cells form Schwann cells that support repair of severed peripheral nerves. Proc Natl Acad Sci U S A, 102:17734–17738.
22.
OhyamaM, TerunumaA, TockCL, RadonovichMF, Pise-MasisonCA, HoppingSB, BradyJN, UdeyMC, VogelJC. 2006. Characterization and isolation of stem cell-enriched human hair follicle bulge cells. J Clin Invest, 116:249–260.
23.
RendlM, LewisL, FuchsE. 2005. Molecular dissection of mesenchymal-epithelial interactions in the hair follicle. PLoS Biol, 3:e331.
24.
WegnerM, StoltCC. 2005. From stem cells to neurons and glia: a Soxist's view of neural development. Trends Neurosci, 28:583–588.
25.
TakahashiK, YamanakaS. 2006. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell, 126:663–676.
26.
TakahashiK, TanabeK, OhnukiM, NaritaM, IchisakaT, TomodaK, YamanakaS. 2007. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell, 131:861–872.
27.
ParkSB, SeoKW, SoAY, SeoMS, YuKR, KangSK, KangKS. 2011. SOX2 has a crucial role in the lineage determination and proliferation of mesenchymal stem cells through Dickkopf-1 and c-MYC. Cell Death Differ, 19:534–545.
28.
ShimJH, KangHH, LeeTR, ShinDW. 2012. Enrichment and characterization of human dermal stem/progenitor cells using collagen, Type IV. J Dermatol Sci, 67:202–205.
29.
ScottMA, NguyenVT, LeviB, JamesAW. 2011. Current methods of adipogenic differentiation of mesenchymal stem cells. Stem Cells Dev, 10:1793–1804.
30.
MontemurroT, AndrioloG, MontelaticiE, WeissmannG, CrisanM, ColnaghiMR, RebullaP, MoscaF, PéaultB, LazzariL. 2011. Differentiation and migration properties of human foetal umbilical cord perivascular cells: potential for lung repair. J Cell Mol Med, 15:796–808.
31.
GaoL, McBeathR, ChenCS. 2010. Stem cell shape regulates a chondrogenic versus myogenic fate through Rac1 and N-cadherin. Stem Cells, 28:564–572.
32.
D'IppolitoG, DiabiraS, HowardGA, MeneiP, RoosBA, SchillerPC. 2004. Marrow-isolated adult multilineage inducible (MIAMI) cells, a unique population of postnatal young and old human cells with extensive expansion and differentiation potential. J Cell Sci, 117:2971–2981.
HughesOR, StewartR, DimmickI, JonesEA. 2009. A critical appraisal of factors affecting the accuracy of results obtained when using flow cytometry in stem cell investigations: where do you put your gates?Cytometry A, 75:803–810.
35.
PochampallyR. 2008. Colony forming unit assays for MSCs. Methods Mol Biol, 449:83–91.
SekiyaI, LarsonBL, SmithJR, PochampallyR, CuiJG, ProckopDJ. 2002. Expansion of human adult stem cells from bone marrow stroma: conditions that maximize the yields of early progenitors and evaluate their quality. Stem Cells, 20:530–541.
38.
JoannidesA, GaughwinP, SchwieningC, MajedH, SterlingJ, CompstonA, ChandranS. 2004. Efficient generation of neural precursors from adult human skin: astrocytes promote neurogenesis from skin-derived stem cells. Lancet, 364:172–178.
39.
ProckopDJ. 1997. Marrow stromal cells as stem cells for nonhematopoietic tissues. Science, 276:71–77.
40.
WeissmanIL. 2000. Stem cells: units of development, units of regeneration, and units in evolution. Cell, 100:157–168.
41.
MezeyE, KeyS, VogelsangG, SzalayovaI, LangeGD, CrainB. 2003. Transplanted bone marrow generates new neurons in human brains. Proc Natl Acad Sci U S A, 100:1364–1369.
42.
LiL, CleversH. 2010. Coexistence of quiescent and active adult stem cells in mammals. Science, 327:542–545.
43.
LysyPA, SmetsF, SibilleC, NajimiM, SokalEM. 2007. Human skin fibroblasts: from mesodermal to hepatocyte-like differentiation. Hepatology, 46:1574–1585.
44.
BiD, ChenFG, ZhangWJ, ZhouGD, CuiL, LiuW, CaoY. 2010. Differentiation of human multipotent dermal fibroblasts into islet-like cell clusters. BMC Cell Biol, 11:46.
45.
ManiniI, GulinoL, GavaB, PierantozziE, CurinaC, RossiD, BrafaA, D'AnielloC, SorrentinoV. 2011. Multi-potent progenitors in freshly isolated and cultured human mesenchymal stem cells: a comparison between adipose and dermal tissue. Cell Tissue Res, 344:85–95.
46.
BezA, CorsiniE, CurtiD, BiggiogeraM, ColomboA, NicosiaRF, PaganoSF, ParatiEA. 2003. Neurosphere and neurosphere-forming cells: morphological and ultrastructural characterization. Brain Res, 993:18–29.
47.
RomanoAC, EspanaEM, YooSH, BudakMT, WolosinJM, TsengSC. 2003. Different cell sizes in human limbal and central corneal basal epithelia measured by confocal microscopy and flow cytometry. Invest Ophthalmol Vis Sci, 44:5125–5129.
48.
MurayamaA, MatsuzakiY, KawaguchiA, ShimazakiT, OkanoH. 2002. Flow cytometric analysis of neural stem cells in the developing and adult mouse brain. J Neurosci Res, 69:837–847.
49.
FauquierT, RizzotiK, DattaniM, Lovell-BadgeR, RobinsonIC. 2008. SOX2-expressing progenitor cells generate all of the major cell types in the adult mouse pituitary gland. Proc Natl Acad Sci U S A, 105:2907–2912.
50.
LiuS, LiuS, WangX, ZhouJ, CaoY, WangF, DuanE. 2011. The PI3K-Akt pathway inhibits senescence and promotes self-renewal of human skin-derived precursors in vitro. Aging Cell, 10:661–674.
YlöstaloJH, BartoshTJ, CobleK, ProckopDJ. 2012. Human mesenchymal stem/stromal cells cultured as spheroids are self-activated to produce prostaglandin E2 that directs stimulated macrophages into an anti-inflammatory phenotype. Stem Cells, 30:2283–2296.
55.
YuKR, YangSR, JungJW, KimH, KoK, HanDW, ParkSB, ChoiSW, KangSK, SchölerH, KangKS. 2012. CD49f enhances multipotency and maintains stemness through the direct regulation of OCT4 and SOX2. Stem Cells, 30:876–887.
56.
RiekstinaU, MucenieceR, CakstinaI, MuiznieksI, AncansJ. 2008. Characterization of human skin-derived mesenchymal stem cell proliferation rate in different growth conditions. Cytotechnology, 58:153–162.
57.
RiekstinaU, CakstinaI, ParfejevsV, HoogduijnM, JankovskisG, MuiznieksI, MucenieceR, AncansJ. 2009. Embryonic stem cell marker expression pattern in human mesenchymal stem cells derived from bone marrow, adipose tissue, heart and dermis. Stem Cell Rev, 5:378–386.
58.
JahodaCA, WhitehouseJ, ReynoldsAJ, HoleN. 2003. Hair follicle dermal cells differentiate into adipogenic and osteogenic lineages. Exp Dermatol, 12:849–859.
59.
HoogduijnMJ, GorjupE, GeneverPG. 2006. Comparative characterization of hair follicle dermal stem cells and bone marrow mesenchymal stem cells. Stem Cells Dev, 15:49–60.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
