Isolation of Mesenchymal Stem Cell from Wharton’s Jelly of Human Umbilical Cord for Application in Wound Healing
Main Article Content
Abstract
Abstract: Mesenchymal stem cell (MSC) is a promising source of novel cell-based therapies, driven by the hope of finding cures for numerous diseases including skin wound healing. In this study, we isolated MSCs from Wharton’s jelly of human umbilical cord by enzymatic method. To determine the effect of MSC conditioned medium on wound healing ability, we examined two MSC conditioned mediums (MSC-CM), which differ in concentration of serum and harvest time. The results showed that in serum starvation condition, MSC-CM showed significantly enhanced keratinocyte migration speed and prolonged culture of MSC in this condition also improve the efficiency of MSC-CM.
Keywords: Wharton’s jelly, Mesenchymal stem cell-conditioned medium, serum starvation, wound healing.References
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Langan TJ, Chou RC. Synchronization of mammalian cell cultures by serum deprivation. Methods Mol Biol., 761 (2011) 75
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[4] Campagnoli C, Roberts IA, Kumar S, Bennett PR, Bellantuono I, Fisk NM, Identification of mesenchymal stem/progenitor cells in human first-trimester fetal blood, liver, and bone marrow, Blood, 98 (2001) 2396.
[5] 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, 22 (2004) 1330.
[6] Kim J, Lee Y, Kim H, Hwang KJ, Kwon HC, Kim SK, Cho DJ, Kang SG, You J, Human amniotic fluid-derived stem cells have characteristics of multipotent stem cells, Cell Prolif, 40 (2007) 75.
[7] De Bari C, Dell'Accio F, Vandenabeele F, Vermeesch JR, Raymackers JM, Luyten FP, Skeletal muscle repair by adult human mesenchymal stem cells from synovial membrane, J Cell Biol, 160 (2003) 909.
[8] Kern S, Eichler H, Stoeve J, Klüter H, Bieback K, Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue, Stem Cells, 24 (2006) 1294.
[9] Mills SJ, Cowin AJ, Kaur P, Pericytes, mesenchymal stem cells and the wound healing process, Cells, 2 (2013) 621.
[10] Jayaraman P, Nathan P, Vasanthan P, Musa S, Govindasamy V, Stem cells conditioned medium: a new approach to skin wound healing management, Cell BiolInt, 37 (2013) 1122.
[11] Paquet-Fifield S, Schlüter H, Li A, Aitken T, Gangatirkar P, Blashki D, Koelmeyer R, Pouliot N, Palatsides M, Ellis S, Brouard N, Zannettino A, Saunders N, Thompson N, Li J, Kaur P, A role for pericytes as microenvironmental regulators of human skin tissue regeneration, J Clin Invest, 119 (2009) 2795.
[12] Chen L, Tredget EE, Wu PY, Wu Y, Paracrine factors of mesenchymal stem cells recruit macrophages and endothelial lineage cells and enhance wound healing, PLoS One, 3 (2008) 1886.
[13] Shohara R, Yamamoto A, Takikawa S, Iwase A, Hibi H, Kikkawa F, Ueda M, Mesenchymal stromal cells of human umbilical cord Wharton's jelly accelerate wound healing by paracrine mechanisms, Cytotherapy, 14 (2012) 1171.
[14] Patrick P, Joshua D, Alaina B, Robert A B, Effect of Serum and Oxygen Concentration on Gene Expression and Secretion of Paracrine Factors by Mesenchymal Stem Cells, International Journal of Cell Biology, 2014 (2014).
[15] Salehinejad P, Alitheen NB, Ali AM, Omar AR, Mohit M, Janzamin E, SamaniFS,Torshizi Z, Nematollahi-Mahani SN, Comparison of different methods for the isolation of mesenchymal stem cells from human umbilical cord Wharton's jelly, In Vitro Cell DevBiolAnim, 48 (2012) 75.
[16] Li M, Zhao Y, Hao H, Dai H, Han Q, Tong C, Liu J, Han W, Fu X, Mesenchymal stem cell-conditioned medium improves the proliferation and migration of keratinocytes in a diabetes-like microenvironment, Int J Low Extrem Wounds, 14 (2015) 73.
[17] Potapova IA, Gaudette GR, Brink PR, Robinson RB, Rosen MR, Cohen IS, Doronin SV, Mesenchymal stem cells support migration, extracellular matrix invasion, proliferation, and survival of endothelial cells in vitro, Stem Cells, 25 (2007) 1761.
[18] Yew TL, Hung YT, Li HY, Chen HW, Chen LL, Tsai KS, Chiou SH, Chao KC, Huang TF, Chen HL, Hung SC, Enhancement of wound healing by human multipotent stromal cell conditioned medium: the paracrine factors and p38 MAPK activation, Cell Transplant, 20 (2011) 693.
[19] Ries C, Egea V, Karow M, Kolb H, Jochum M, Neth P. MMP-2, MT1-MMP, and TIMP-2 are essential for the invasive capacity of human mesenchymal stem cells: differential regulation by inflammatory cytokines, Blood, 109 (2007) 4055.
Langan TJ, Chou RC. Synchronization of mammalian cell cultures by serum deprivation. Methods Mol Biol., 761 (2011) 75