Isolation and characterization of in vitro culture of hair follicle cells differentiated from umbilical cord blood mesenchymal stem cells

Bu,Zhang‑Yu; Wu,Li‑Min; Yu,Xiao‑Hong; Zhong,Jian‑Bo; Yang,Ping; Chen,Jian

doi:10.3892/etm.2017.4456

Isolation and characterization of in vitro culture of hair follicle cells differentiated from umbilical cord blood mesenchymal stem cells

Authors:
- Zhang‑Yu Bu
- Li‑Min Wu
- Xiao‑Hong Yu
- Jian‑Bo Zhong
- Ping Yang
- Jian Chen
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Affiliations: Department of Dermatology, Hangzhou First People's Hospital, Nanjing Medical University, Hangzhou, Zhejiang 310006, P.R. China
Published online on: May 16, 2017 https://doi.org/10.3892/etm.2017.4456
Pages: 303-307
Copyright: © Bu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present investigation explored the in vitro culture, isolation and characterization of hair follicle cell differentiation from umbilical cord blood mesenchymal stem cells (MSCs). Flow cytometry was used to obtain MSCs from the isolation and purification of human umbilical cord blood MSCs. Culture suspension of hair follicle organ was centrifuged and the supernatant used in the culture medium of MSCs, and the entire process of induced differentiation was recorded by photomicroscopy. The expression level of surface marker CK15 of hair follicle cells obtained from induced differentiation was detected with immunofluorescence. RT‑PCR method was used to further detect the difference in expression of CK15 between hair follicle cells and umbilical cord blood MSCs, and statistical analysis was carried out. CD44+CD29+ double‑labeled cells accounted for 50.8% of all the samples of umbilical cord blood MSCs in this study. The diameter of hair follicle cells differentiated from umbilical cord blood stem cells reached 800x10‑3 mm after 3 weeks of cell culture. Based on the detection and colocalization of CK15 expression in induced hair follicle cells, the overlap ratio between CK15 and nuclei reached 83% in hair follicle cells, which was obviously higher than that in umbilical cord blood stem cells. The difference had statistical significance (P<0.05). In conclusion, hair follicle cells can be successfully differentiated from umbilical cord blood stem cells by using the supernatant from hair follicle cells. This method can be used for high‑speed induced differentiation with high purity, which is promising for clinical application.

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1	Codinach M, Blanco M, Ortega I, Lloret M, Reales L, Coca MI, Torrents S, Doral M, Oliver-Vila I, Requena-Montero M, et al: Design and validation of a consistent and reproducible manufacture process for the production of clinical-grade bone marrow-derived multipotent mesenchymal stromal cells. Cytotherapy. 18:1197–1208. 2016. View Article : Google Scholar : PubMed/NCBI
2	Kumar K, Agarwal P, Das K, Mili B, Madhusoodan AP, Kumar A and Bag S: Isolation and characterization of mesenchymal stem cells from caprine umbilical cord tissue matrix. Tissue Cell. 48:653–658. 2016. View Article : Google Scholar : PubMed/NCBI
3	Del Valle-Echevarria AR, Sanseverino W, Garcia-Mas J and Havey MJ: Pentatricopeptide repeat 336 as the candidate gene for paternal sorting of mitochondria (Psm) in cucumber. Theor Appl Genet. 129:1951–1959. 2016. View Article : Google Scholar : PubMed/NCBI
4	de Moor JS, Dowling EC, Ekwueme DU, Guy GP Jr, Rodriguez J, Virgo KS, Han X, Kent EE, Li C, Litzelman K, et al: Employment implications of informal cancer caregiving. J Cancer Surviv. 11:48–57. 2017. View Article : Google Scholar : PubMed/NCBI
5	Sigurjónsson OE, Guðmundsson KO and Guðmundsson S: Mesenchymal stem cells. A review. Laeknabladid. 87:627–632. 2001.(In Icelandic). PubMed/NCBI
6	Maranda EL, Rodriguez-Menocal L and Badiavas EV: Role of mesenchymal stem cells in dermal repair in burns and diabetic wounds. Curr Stem Cell Res Ther. 12:61–70. 2017. View Article : Google Scholar : PubMed/NCBI
7	Astori G, Amati E, Bambi F, Bernardi M, Chieregato K, Schäfer R, Sella S and Rodeghiero F: Platelet lysate as a substitute for animal serum for the ex-vivo expansion of mesenchymal stem/stromal cells: Present and future. Stem Cell Res Ther. 7:93–100. 2016. View Article : Google Scholar : PubMed/NCBI
8	Dyrna F, Herbst E, Hoberman A, Imhoff AB and Schmitt A: Stem cell procedures in arthroscopic surgery. Eur J Med Res. 21:29–36. 2016. View Article : Google Scholar : PubMed/NCBI
9	Jiang LH, Hao Y, Mousawi F, Peng H and Yang X: Expression of P2 purinergic receptors in mesenchymal stem cells and their roles in extracellular nucleotide regulation of cell functions. J Cell Physiol. 232:287–297. 2017. View Article : Google Scholar : PubMed/NCBI
10	Morris AD, Chen J, Lau E and Poh J: Domperidone-associated QT interval prolongation in non-oncologic pediatric patients: A review of the literature. Can J Hosp Pharm. 69:224–230. 2016.PubMed/NCBI
11	Kavosi Z, Khorrami M Sarikhani, Keshavarz K, Jafari A, Meshkini A Hashemi, Safaei HR and Nikfar S: Is taurolidine-citrate an effective and cost-effective hemodialysis catheter lock solution? A systematic review and cost-effectiveness analysis. Med J Islam Repub Iran. 30:347–358. 2016.PubMed/NCBI
12	Shi X, Lv S, He X, Liu X, Sun M, Li M, Chi G and Li Y: Differentiation of hepatocytes from induced pluripotent stem cells derived from human hair follicle mesenchymal stem cells. Cell Tissue Res. 366:89–99. 2016. View Article : Google Scholar : PubMed/NCBI
13	Maruyama CL, Leigh NJ, Nelson JW, McCall AD, Mellas RE, Lei P, Andreadis ST and Baker OJ: Stem cell-soluble signals enhance multilumen formation in SMG cell clusters. J Dent Res. 94:1610–1617. 2015. View Article : Google Scholar : PubMed/NCBI
14	Son S, Liang MS, Lei P, Xue X, Furlani EP and Andreadis ST: Magnetofection mediated transient NANOG overexpression enhances proliferation and myogenic differentiation of human hair follicle derived mesenchymal stem cells. Bioconjug Chem. 26:1314–1327. 2015. View Article : Google Scholar : PubMed/NCBI
15	Maleki M, Ghanbarvand F, Behvarz M Reza, Ejtemaei M and Ghadirkhomi E: Comparison of mesenchymal stem cell markers in multiple human adult stem cells. Int J Stem Cells. 7:118–126. 2014. View Article : Google Scholar : PubMed/NCBI
16	Dong L, Hao H, Xia L, Liu J, Ti D, Tong C, Hou Q, Han Q, Zhao Y, Liu H, et al: Treatment of MSCs with Wnt1a-conditioned medium activates DP cells and promotes hair follicle regrowth. Sci Rep. 4:5432–5440. 2014. View Article : Google Scholar : PubMed/NCBI
17	Wu M, Guo X, Yang L, Wang Y, Tang Y, Yang Y and Liu H: Mesenchymal stem cells with modification of junctional adhesion molecule a induce hair formation. Stem Cells Transl Med. 3:481–488. 2014. View Article : Google Scholar : PubMed/NCBI
18	Wang Y, Liu J, Tan X, Li G, Gao Y, Liu X, Zhang L and Li Y: Induced pluripotent stem cells from human hair follicle mesenchymal stem cells. Stem Cell Rev. 9:451–460. 2013. View Article : Google Scholar : PubMed/NCBI
19	Gola M, Czajkowski R, Bajek A, Dura A and Drewa T: Melanocyte stem cells: Biology and current aspects. Med Sci Monit. 18:RA155–RA159. 2012. View Article : Google Scholar : PubMed/NCBI
20	DiDomenico L, Landsman AR, Emch KJ and Landsman A: A prospective comparison of diabetic foot ulcers treated with either a cryopreserved skin allograft or a bioengineered skin substitute. Wounds. 23:184–189. 2011.PubMed/NCBI
21	Liang MS and Andreadis ST: Engineering fibrin-binding TGF-β1 for sustained signaling and contractile function of MSC based vascular constructs. Biomaterials. 32:8684–8693. 2011. View Article : Google Scholar : PubMed/NCBI