MEF/KSF‑conditioned culture medium: An effective method for <em>in vitro</em> culture of mouse dermal papilla cells with osteogenic differentiation potential

Xu,Liang; Gao,Wenlan; Bai,Shanshan; Duan,Huichuan; Pan,Xiaogang; Wu,Wei

doi:10.3892/etm.2021.10260

MEF/KSF‑conditioned culture medium: An effective method for in vitro culture of mouse dermal papilla cells with osteogenic differentiation potential

Authors:
- Liang Xu
- Wenlan Gao
- Shanshan Bai
- Huichuan Duan
- Xiaogang Pan
- Wei Wu
View Affiliations

Affiliations: Department of Plastic and Reconstructive Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, P.R. China, Department of Stomatology, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, P.R. China, Department of Orthodontics, Shanghai Ninth Peoples' Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, P.R. China
Published online on: June 3, 2021 https://doi.org/10.3892/etm.2021.10260
Article Number: 828
Copyright: © Xu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Hair follicle stem cells are pluripotent and have a self‑renewal capacity and multi‑differentiation potential in vitro. As hair follicle stem cells can be easily sampled from the skin and hair of clinical patients at a considerable quantity, these cells have potential applications in wound repair and skin tissue engineering. Effective approaches for the in vitro culture and amplification of mouse hair follicle stem cells, as well as the in vitro osteogenic differentiation potential and cell source when obtaining mouse‑separated cells were examined. Serial subculture was performed in different culture systems. Cell source was detected based on the relevant surface markers derived from mouse hair follicles at the gene and protein levels, and the differential potential was determined. The proliferative ability of hair follicle‑derived stem cells obtained from mouse embryonic fibroblast (MEF)/keratinocyte serum‑free medium (KSF)‑conditioned medium was the highest among all culture systems. The induced group had a stronger osteogenic differentiation potential compared with the non‑induced group, indicating that the cells obtained from MEF/KSF‑conditioned medium were cells derived from the hair follicle dermal papilla. Therefore, the strong osteogenic differentiation potential of the hair follicle‑derived mesenchymal stem cells was screened with MEF/KSF‑conditioned culture medium following amplification, and biological characteristics similar to those of hair follicle dermal papilla cells were observed.

View Figures

View References

1	Amoh Y, Kanoh M, Niiyama S, Hamada Y, Kawahara K, Sato Y, Hoffman RM and Katsuoka K: Human hair follicle pluripotent stem (hfPS) cells promote regeneration of peripheral-nerve injury: An advantageous alternative to ES and iPS cells. J Cell Biochem. 107:1016–1020. 2009.PubMed/NCBI View Article : Google Scholar
2	Carrasco E, Calvo MI, Blázquez-Castro A, Vecchio D, Zamarrón A, de Almeida IJD, Stockert JC, Hamblin MR, Juarranz Á and Espada J: Photoactivation of ROS production in situ transiently activates cell proliferation in mouse skin and in the hair follicle stem cell niche promoting hair growth and wound healing. J Invest Dermatol. 135:2611–2622. 2015.PubMed/NCBI View Article : Google Scholar
3	Wang B, Liu XM, Liu ZN, Wang Y, Han X, Lian AB, Mu Y, Jin MH and Liu JY: Human hair follicle-derived mesenchymal stem cells: Isolation, expansion, and differentiation. World J Stem Cells. 12:462–470. 2020.PubMed/NCBI View Article : Google Scholar
4	Castro AR and Logarinho E: Tissue engineering strategies for human hair follicle regeneration: How far from a hairy goal. Stem Cells Transl Med. 9:342–350. 2020.PubMed/NCBI View Article : Google Scholar
5	Li B, Hu W, Ma K, Zhang C and Fu X: Are hair follicle stem cells promising candidates for wound healing? Expert Opin Biol Ther. 19:119–128. 2019.PubMed/NCBI View Article : Google Scholar
6	Brockmann I, Ehrenpfordt J, Sturmheit T, Brandenburger M, Kruse C, Zille M, Rose D and Boltze J: Skin-derived stem cells for wound treatment using cultured epidermal autografts: Clinical applications and challenges. Stem Cells Int. 2018(4623615)2018.PubMed/NCBI View Article : Google Scholar
7	Bergeron L, Busuttil V and Botto JM: Multipotentiality of skin-derived precursors: Application to the regeneration of skin and other tissues. Int J Cosmet Sci. 42:5–15. 2020.PubMed/NCBI View Article : Google Scholar
8	Martínez ML, Escario E, Poblet E, Sánchez D, Buchón FF, Izeta A and Jimenez F: Hair follicle-containing punch grafts accelerate chronic ulcer healing: A randomized controlled trial. J Am Acad Dermatol. 75:1007–1014. 2016.PubMed/NCBI View Article : Google Scholar
9	Jahoda CA, Whitehouse J, Reynolds AJ and Hole N: Hair follicle dermal cells differentiate into adipogenic and osteogenic lineages. Exp Dermatol. 12:849–859. 2003.PubMed/NCBI View Article : Google Scholar
10	Tögel F and Westenfelder C: Adult bone marrow-derived stem cells for organ regeneration and repair. Dev Dyn. 236:3321–3331. 2007.PubMed/NCBI View Article : Google Scholar
11	Conejero JA, Lee JA, Parrett BM, Terry M, Wear-Maggitti K, Grant RT and Breitbart AS: Repair of palatal bone defects using osteogenically differentiated fat-derived stem cells. Plast Reconstr Surg. 117:857–863. 2006.PubMed/NCBI View Article : Google Scholar
12	Bueno DF, Kerkis I, Costa AM, Martins MT, Kobayashi GS, Zucconi E, Fanganiello RD, Salles FT, Almeida AB, do Amaral CE, et al: New source of muscle-derived stem cells with potential for alveolar bone reconstruction in cleft lip and/or palate patients. Tissue Eng Part A. 15:427–435. 2009.PubMed/NCBI View Article : Google Scholar
13	Hoogduijn MJ, Gorjup E and Genever PG: Comparative characterization of hair follicle dermal stem cells and bone marrow mesenchymal stem cells. Stem Cells Dev. 15:49–60. 2006.PubMed/NCBI View Article : Google Scholar
14	Schneider MR, Schmidt-Ullrich R and Paus R: The hair follicle as a dynamic miniorgan. Curr Biol. 19:R132–R142. 2009.PubMed/NCBI View Article : Google Scholar
15	Rochat A, Kobayashi K and Barrandon Y: Location of stem cells of human hair follicles by clonal analysis. Cell. 76:1063–1073. 1994.PubMed/NCBI View Article : Google Scholar
16	Lin BJ, Lin GY, Zhu JY, Yin GQ, Huang D and Yan YY: LncRNA-PCAT1 maintains characteristics of dermal papilla cells and promotes hair follicle regeneration by regulating miR-329/Wnt10b axis. Exp Cell Res. 24(112031)2020.PubMed/NCBI View Article : Google Scholar
17	Ohyama M: Hair follicle bulge: A fascinating reservoir of epithelial stem cells. J Dermatol Sci. 46:81–89. 2007.PubMed/NCBI View Article : Google Scholar
18	Cotsarelis G: Gene expression profiling gets to the root of human hair follicle stem cells. J Clin Invest. 116:19–22. 2006.PubMed/NCBI View Article : Google Scholar
19	Moll I: Differential epithelial outgrowth of plucked and microdissected human hair follicles in explant culture. Arch Dermatol Res. 288:604–610. 1996.PubMed/NCBI View Article : Google Scholar
20	Chen L, Duan H, Xie F, Gao Z, Wu X, Chen F and Wu W: Tetrahydroxystilbene glucoside effectively prevents apoptosis induced hair loss. Biomed Res Int. 2018(1380146)2008.PubMed/NCBI View Article : Google Scholar
21	Chi W, Wu E and Morgan BA: Dermal papilla cell number specifies hair size, shape and cycling and its reduction causes follicular decline. Development. 140:1676–1683. 2013.PubMed/NCBI View Article : Google Scholar
22	Lammens J, Maréchal M, Delport H, Geris L, Oppermann H, Vukicevic S and Luyten FP: A cell-based combination product for the repair of large bone defects. Bone. 138(115511)2020.PubMed/NCBI View Article : Google Scholar
23	Vogler HW: Basic bioengineering concepts, Concepts in bone performance, failure, and osteosynthesis. Clin Podiatry. 2:161–189. 1985.PubMed/NCBI
24	Kneser U, Schaefer DJ, Polykandriotis E and Horch RE: Tissue enginering of bone: The reconstructive surgeon^'s point of view. J Cell Mol Med. 10:7–19. 2006.PubMed/NCBI View Article : Google Scholar
25	Li X, Yuan Z, Wei X, Li H, Zhao G, Miao J, Wu D, Liu B, Cao S, An D, et al: Application potential of bone marrow mesenchymal stem cell (BMSCs) based tissue-engineering for spinal cord defect repair in rat fetuses with spina bifida aperta. J Mater Sci Mater Med. 27(77)2016.PubMed/NCBI View Article : Google Scholar
26	Loukogeorgakis S and De Coppi P: Stem cells form amniotic fluid-potential for regenerative medicine. Best Pract Clin Obstet Gynaecol. 31:45–57. 2015.PubMed/NCBI View Article : Google Scholar
27	Sypecka J and Sarnowska A: Mesenchymal cells of umbilical cord and umbilical cord blood as a source of human oligodendrocyte progenitors. Life Sci. 139:24–29. 2015.PubMed/NCBI View Article : Google Scholar
28	Jin SE and Sung JH: Hair regeneration using adipose-derived stem cells. Histol Histopathol. 31:249–256. 2016.PubMed/NCBI View Article : Google Scholar
29	Cheng CS, Davis BN, Madden L, Bursac N and Truskey GA: Physiology and metabolism of tissue-engineered skeletal muscle. Exp Biol Med (Maywood). 239:1203–1214. 2014.PubMed/NCBI View Article : Google Scholar
30	Tatullo M, Marrelli M, Shakesheff KM and White LJ: Dental pulp stem cells: Function, isolation and applications in regenerative medicine. J Tissue Eng Regen Med. 9:1205–1216. 2015.PubMed/NCBI View Article : Google Scholar
31	Cotsarelis G, Sun TT and Lavker RM: Label-retaining cells reside in the bulge area of pilosebaceous unit: Implication for follicular stem cell, hair cycle, and skin carcinogenesis. Cell. 61:1329–1337. 1990.PubMed/NCBI View Article : Google Scholar
32	Taylor G, Lehrer MS, Jensen PJ, Sun TT and Lavker RM: Involvement of follicular stem cell in forming not only the follicle but also the epidermis. Cell. 102:451–461. 2000.PubMed/NCBI View Article : Google Scholar
33	Kurata S, Itami S, Terashi H and Takayasu S: Successful transplantation of cultured human outer root sheath cells as epithelium. Ann Plast Surg. 33:290–294. 1994.PubMed/NCBI View Article : Google Scholar
34	Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS and Jones JM: Embryonic stem cell lines derived from human blastocysts. Science. 282:1145–1147. 1998.PubMed/NCBI View Article : Google Scholar
35	Hu J, Hu S, Ma Q, Wang X, Zhou Z, Zhang W, Sun X, Zhu W, Qian H and Xu W: Immortalized mouse fetal liver stromal cells support growth and maintenance of human embryonic stem cells. Oncol Rep. 28:1385–1391. 2012.PubMed/NCBI View Article : Google Scholar
36	Chin AC, Fong WJ, Goh LT, Philp R, Oh SK and Choo AB: Identification of proteins from feeder conditioned medium that support human embryonic stem cells. J Biotechnol. 130:320–328. 2007.PubMed/NCBI View Article : Google Scholar
37	Shwartz Y, Gonzalez-Celeiro M, Chen CL, Pasolli HA, Sheu SH, Fan SMY, Shamsi F, Assaad S, Lin ETY, Zhang B, et al: Cell types promoting goosebumps form a niche to regulate hair follicle stem cells. Cell. 182:578–593. 2020.PubMed/NCBI View Article : Google Scholar
38	Bajpai VK, Mistriotis P and Andreadis ST: Clonal multipotency and effect of long-term in vitro expansion on differentiation potential of human hair follicle derived mesenchymal stem cells. Stem Cell Res. 8:74–84. 2012.PubMed/NCBI View Article : Google Scholar
39	Morris RJ, Liu Y, Marles L, Yang Z, Trempus C, Li S, Lin JS, Sawicki JA and Cotsarelis G: Capturing and profiling adult hair follicle stem cells. Nat Biotechnol. 22:411–417. 2004.PubMed/NCBI View Article : Google Scholar
40	Morgan BA: The dermal papilla: An instruction niche for epithelial stem and progenitor cells in development and regeneration of the hair follicle. Cold Spring Harb Perspect Med. 4(a015180)2014.PubMed/NCBI View Article : Google Scholar
41	McElwee KJ, Kissling S, Wenzel E, Huth A and Hoffmann R: Cultured peribulbar dermal sheath cells can induce hair follicle development and contribute to the dermal sheath and dermal papilla. J Invest Dermatol. 121:1267–1275. 2003.PubMed/NCBI View Article : Google Scholar
42	Liu JY, Peng HF, Gopinath S, Tian J and Andreadis ST: Derivation of functional smooth muscle cells from multipotent human hair follicle mesenchymal stem cells. Tissue Eng Part A. 16:2553–2564. 2010.PubMed/NCBI View Article : Google Scholar
43	Yu H, Kumar SM, Kossenkov AV, Showe L and Xu X: Stem cells with neural crest characteristics derived from the bulge region of cultured human hair follicles. J Invest Dermatol. 130:1227–1236. 2010.PubMed/NCBI View Article : Google Scholar
44	Boden SD: The ABCs of BMPs. Orthop Nurs. 24:49–52; quiz53-54. 2005.PubMed/NCBI View Article : Google Scholar
45	Lin CM, Yuan YP, Chen XC, Li HH, Cai BZ, Liu Y, Zhang H, Li Y and Huang K: Expression of Wnt/β-catenin signaling, stem-cell markers and proliferating cell markers in rat whisker hair follicles. J Mol Histol. 46:233–240. 2015.PubMed/NCBI View Article : Google Scholar
46	Li F, Song N, Tombran-Tink J and Niyibizi C: Pigment epithelium-derived factor enhances differentiation and mineral deposition of human mesenchymal stem cells. Stem Cells. 12:2714–2723. 2013.PubMed/NCBI View Article : Google Scholar
47	Lyles JM, Amin W, Bock E and Weill CL: Regulation of NCAM by growth factors in serum-free myotube cultures. J Neurosci Res. 34:273–286. 1993.PubMed/NCBI View Article : Google Scholar
48	Seo E, Basu-Roy U, Zavadil J, Basilico C and Mansukhani A: Distinct functions of sox2 control self-renewal and differentiation in the osteoblast lineage. Mol Cell Biol. 22:4593–4608. 2011.PubMed/NCBI View Article : Google Scholar
49	O'Connor MD, Kardel MD, Iosfina I, Youssef D, Lu M, Li MM, Vercauteren S, Nagy A and Eaves CJ: Alkaline phosphatase-positive colony formation is a sensitive, specific, and quantitative indicator of undifferentiated human embryonic stem cells. Stem Cells. 5:1109–1116. 2008.PubMed/NCBI View Article : Google Scholar