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Frontiers of Medicine

ISSN 2095-0217

ISSN 2095-0225(Online)

CN 11-5983/R

Postal Subscription Code 80-967

2018 Impact Factor: 1.847

Front Med    2013, Vol. 7 Issue (3) : 345-353    https://doi.org/10.1007/s11684-013-0282-2
RESEARCH ARTICLE
Capacity of human umbilical cord-derived mesenchymal stem cells to differentiate into sweat gland-like cells: a preclinical study
Siming Yang1,2, Kui Ma2, Changjiang Feng2, Yan Wu2, Yao Wang2, Sha Huang1,2, Xiaobing Fu1,2()
1. Institute of Basic Medical Sciences, PLA General Hospital, the PLA Medical College, Beijing 100853, China; 2. Burns Institute, the First Affiliated Hospital, PLA General Hospital, Beijing 100048, China
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Abstract

Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) possess various advantageous properties, including self-renewal, extended proliferation potential, multi-lineage differentiation potential and capacity for differentiating into sweat gland-like cells in certain conditions. However, little is known about the effect of clinical-grade culture conditions on these properties and on the differentiative potential of hUC-MSCs. In this study, we sought to investigate the properties of hUC-MSCs expanded with animal serum free culture media (ASFCM) in order to determine their potential for differentiation into sweat gland-like cells. We found that primary cultures of hUC-MSCs could be established with ASFCM. Moreover, cells cultured in ASFCM showed vigorous proliferation comparable to those of cells grown in classical culture conditions containing fetal bovine serum (FBS). Morphology of hUC-MSCs cultured in ASFCM was comparable to those of cells grown under classical culture conditions, and hUC-MSCs grown in both of the two culture conditions tested showed the typical antigen profile of MSCs—positive for CD29, CD44, CD90, and CD105, and negative for CD34 and CD45, as expected. Chromosomal aberration assay revealed that the cells were stable after long-term culture under both culture conditions. Like normal cultured MSCs, hUC-MSCs induced under ASFCM conditions exhibited expression of the same markers (CEA, CK14 and CK19) and developmental genes (EDA and EDAR) that are characteristic of normal sweat gland cells. Taken together, our findings indicate that the classical culture medium used to differentiate hUC-MSCs into sweat gland-like cells can be replaced safely by ASFCM for clinical purposes.

Keywords umbilical cord      mesenchymal stem cells      sweat gland      preclinical     
Corresponding Author(s): Fu Xiaobing,Email:fuxb@cgw.net.cn; fuxiaobing@vip.sina.com   
Issue Date: 05 September 2013
 Cite this article:   
Siming Yang,Kui Ma,Changjiang Feng, et al. Capacity of human umbilical cord-derived mesenchymal stem cells to differentiate into sweat gland-like cells: a preclinical study[J]. Front Med, 2013, 7(3): 345-353.
 URL:  
https://academic.hep.com.cn/fmd/EN/10.1007/s11684-013-0282-2
https://academic.hep.com.cn/fmd/EN/Y2013/V7/I3/345
Fig.1  Panel I: Morphology of hUC-MSCs of two culture groups. (A) Cells of group 1 at passage 1; (B) Cells of group 2 at passage 2; (C) Cells of group 1 at passage 3; (D) Cells of group 2 at passage 3; = 3 for each group; scale bar= 50 μm. Panel II: Expression of surface markers by flow cytometry analysis. The mean percentages of each phenotype were compared between group 1 and group 2. Both the positive percentages of hUC-MSCs for mesenchymal antigens (CD29, CD44, CD90, and CD105) and the mean negative percentages for hematopoietic stem cells (CD34, CD45) showed no significant difference between group 1 and group 2. Markers of sweat gland cells (CEA, CK14) were all negative in group 1 and group 2 ( = 3 for each group).
Fig.2  Proliferative potency of hUC-MSCs in different culture groups determined by MTT analysis (data shown as mean±SEM). Induction groups 1 and 2 represent previous culture conditions groups 1 and 2, respectively; control group represent cells cultured in basic medium. Proliferation of hUC-MSCs in both induction groups was distinctively lower than that in the control group. (*<0.05, = 3 for each group). No distinct difference in the proliferative potency was observed between induction group 1 and group 2.
Fig.3  Morphology of hUC-MSCs in different conditioned induction media. (A) Cells of induction group 1 were cultured for 1 week and their morphology changed gradually; (B) Cells of induction group 2 were also cultured for 1 week and also changed gradually; (C) Cells of induction group 1 differentiated into sweat gland-like cells after 3 weeks of induction; (D) Cells of induction group 2 differentiated into sweat gland-like cells after 3 weeks of induction. ( = 3 for each group; bar= 50 μm.)
Fig.4  Expression of CEA, CK14 and CK19 in different induction groups (mean±SEM) after 3 weeks. The mean expression percentages of sweat gland markers between induction group 1 and 2 showed no significant difference; >0.05, unpaired post-Bonferroni -test; = 5 for each group.
Fig.5  Expression of EDA and EDAR in different induction groups and normal SGCs. (A) RT-PCR analysis showed gene expression profiles (EDA and EDAR) of sweat gland-like cells relative to uninduced hUC-MSCs and normal sweat gland cells. (B) Western blotting analysis was conducted to assess the expression level of EDA and EDAR in each group. (C) Greyscale analysis of RT-PCR bands; expression of EDA and EDAR between groups 1 and 2 revealed no significant difference; >0.05, unpaired post-Bonferroni -test; = 3 for each group. hUC-MSCs were taken as negative control, while normal SGCs were taken as positive control. SGCs, sweat gland cells; hUC-MSCs, human umbilical cord Wharton’s jelly-derived mesenchymal stem cells; EDA, anhidrotic ectodermal dysplasia; EDAR, anhidrotic ectodermal dysplasia receptor.
Fig.6  Karyotype analysis showed no chromosomal alteration. (A) The 10th passage cells in induction group 1; (B)The 10th passage cells in induction group 2.
1 Yang S, Huang S, Feng C, Fu X. Umbilical cord-derived mesenchymal stem cells: strategies, challenges, and potential for cutaneous regeneration. Front Med 2012; 6(1): 41–47
doi: 10.1007/s11684-012-0175-9 pmid:22460447
2 Lu LL, Liu YJ, Yang SG, Zhao QJ, Wang X, Gong W, Han ZB, Xu ZS, Lu YX, Liu D, Chen ZZ, Han ZC. Isolation and characterization of human umbilical cord mesenchymal stem cells with hematopoiesis-supportive function and other potentials. Haematologica 2006; 91(8): 1017–1026
pmid:16870554
3 Kim JW, Kim SY, Park SY, Kim YM, Kim JM, Lee MH, Ryu HM. Mesenchymal progenitor cells in the human umbilical cord. Ann Hematol 2004; 83(12): 733–738
doi: 10.1007/s00277-004-0918-z pmid:15372203
4 Wang HS, Hung SC, Peng ST, Huang CC, Wei HM, Guo YJ, Fu YS, Lai MC, Chen CC. Mesenchymal stem cells in the Wharton’s jelly of the human umbilical cord. Stem Cells 2004; 22(7): 1330–1337
doi: 10.1634/stemcells.2004-0013 pmid:15579650
5 Friedenstein AJ, Gorskaja JF, Kulagina NN. Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp Hematol 1976; 4(5): 267–274
pmid:976387
6 Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR. Multilineage potential of adult human mesenchymal stem cells. Science 1999; 284(5411): 143–147
doi: 10.1126/science.284.5411.143 pmid:10102814
7 Lennon DP, Edmison JM, Caplan AI. Cultivation of rat marrow-derived mesenchymal stem cells in reduced oxygen tension: effects on in vitro and in vivo osteochondrogenesis. J Cell Physiol 2001; 187(3): 345–355
doi: 10.1002/jcp.1081 pmid:11319758
8 Sekiya I, Vuoristo JT, Larson BL, Prockop DJ. In vitro cartilage formation by human adult stem cells from bone marrow stroma defines the sequence of cellular and molecular events during chondrogenesis. Proc Natl Acad Sci USA 2002; 99(7): 4397–4402
doi: 10.1073/pnas.052716199 pmid:11917104
9 Fu YS, Cheng YC, Lin MY, Cheng H, Chu PM, Chou SC, Shih YH, Ko MH, Sung MS. Conversion of human umbilical cord mesenchymal stem cells in Wharton’s jelly to dopaminergic neurons in vitro: potential therapeutic application for Parkinsonism. Stem Cells 2006; 24(1): 115–124
doi: 10.1634/stemcells.2005-0053 pmid:16099997
10 Wu KH, Zhou B, Lu SH, Feng B, Yang SG, Du WT, Gu DS, Han ZC, Liu YL. In vitro and in vivo differentiation of human umbilical cord derived stem cells into endothelial cells. J Cell Biochem 2007; 100(3): 608–616
doi: 10.1002/jcb.21078 pmid:16960877
11 Weiss ML, Anderson C, Medicetty S, Seshareddy KB, Weiss RJ, VanderWerff I, Troyer D, McIntosh KR. Immune properties of human umbilical cord Wharton’s jelly-derived cells. Stem Cells 2008; 26(11): 2865–2874
doi: 10.1634/stemcells.2007-1028 pmid:18703664
12 Sheng ZY, Fu XB, Cai S, Lei YH, Sun TZ, Bai XD, Chen ML. Functional sweat gland implantation: a report of two cases. Med J Chin PLA (Jie Fang Jun Yi Xue Za Zhi) . 2008; 33(4): 363–368 (in Chinese)
13 Sheng Z, Fu X, Cai S, Lei Y, Sun T, Bai X, Chen M. Regeneration of functional sweat gland-like structures by transplanted differentiated bone marrow mesenchymal stem cells. Wound Repair Regen 2009; 17(3): 427–435
doi: 10.1111/j.1524-475X.2009.00474.x pmid:19660052
14 Xu Y, Huang S, Ma K, Fu X, Han W, Sheng Z. Promising new potential for mesenchymal stem cells derived from human umbilical cord Wharton’s jelly: sweat gland cell-like differentiative capacity. J Tissue Eng Regen Med 2012; 6(8): 645–654
doi: 10.1002/term.468 pmid:21916019
15 Huang P, Lin LM, Wu XY, Tang QL, Feng XY, Lin GY, Lin X, Wang HW, Huang TH, Ma L. Differentiation of human umbilical cord Wharton’s jelly-derived mesenchymal stem cells into germ-like cells in vitro. J Cell Biochem 2010; 109(4): 747–754
pmid:20052672
16 Mikkola ML. TNF superfamily in skin appendage development. Cytokine Growth Factor Rev 2008; 19(3-4): 219–230
doi: 10.1016/j.cytogfr.2008.04.008 pmid:18495521
17 Rao MS, Mattson MP. Stem cells and aging: expanding the possibilities. Mech Ageing Dev 2001; 122(7): 713–734
doi: 10.1016/S0047-6374(01)00224-X pmid:11322994
18 Saga K. Structure and function of human sweat glands studied with histochemistry and cytochemistry. Prog Histochem Cytochem 2002; 37(4): 323–386
doi: 10.1016/S0079-6336(02)80005-5 pmid:12365351
19 Biedermann T, Pontiggia L, B?ttcher-Haberzeth S, Tharakan S, Braziulis E, Schiestl C, Meuli M, Reichmann E. Human eccrine sweat gland cells can reconstitute a stratified epidermis. J Invest Dermatol 2010; 130(8): 1996–2009
doi: 10.1038/jid.2010.83 pmid:20376062
20 Ball LM, Bernardo ME, Roelofs H, Lankester A, Cometa A, Egeler RM, Locatelli F, Fibbe WE. Cotransplantation of ex vivo expanded mesenchymal stem cells accelerates lymphocyte recovery and may reduce the risk of graft failure in haploidentical hematopoietic stem-cell transplantation. Blood 2007; 110(7): 2764–2767
doi: 10.1182/blood-2007-04-087056 pmid:17638847
21 Le Blanc K, Frassoni F, Ball L, Locatelli F, Roelofs H, Lewis I, Lanino E, Sundberg B, Bernardo ME, Remberger M, Dini G, Egeler RM, Bacigalupo A, Fibbe W, Ringdén O; Developmental Committee of the European Group for Blood and Marrow Transplantation. Mesenchymal stem cells for treatment of steroid-resistant, severe, acute graft-versus-host disease: a phase II study. Lancet 2008; 371(9624): 1579–1586
doi: 10.1016/S0140-6736(08)60690-X pmid:18468541
22 Christopeit M, Schendel M, F?ll J, Müller LP, Keysser G, Behre G. Marked improvement of severe progressive systemic sclerosis after transplantation of mesenchymal stem cells from an allogeneic haploidentical-related donor mediated by ligation of CD137L. Leukemia 2008; 22(5): 1062–1064
doi: 10.1038/sj.leu.2404996 pmid:17972956
23 Cui CY, Schlessinger D. EDA signaling and skin appendage development. Cell Cycle 2006; 5(21): 2477–2483
doi: 10.4161/cc.5.21.3403 pmid:17102627
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