Acta Medica Iranica 2018. 56(1):28-33.

Wharton's Jelly Derived-Mesenchymal Stem Cells: Isolation and Characterization
Hossein Ranjbaran, Saeid Abediankenari, Masoud Mohammadi, Narjes Jafari, Alireza Khalilian, Zahra Rahmani, Marzei Momeninezhad Amiri, Pirooz Ebrahimi


Wharton`s jelly-derived mesenchymal stem cells (WJ-MSCs), have a high proliferation valency and they do not produce teratogen or carcinogen after subsequent transplantation. They are known as regenerative medicine. Thus more research is needed on the isolation and characterization of mesenchymal stem cells. In this experimental study, we obtained Wharton's jelly tissues from mothers during normal vaginal delivery, after obtaining their informed consent. Mesenchymal stem cells were isolated from cultured Wharton`s jelly, cultured, and were then examined for their proliferation, immunophenotypes, and differentiation capacities. The immunophenotypes of WJ-MSCs were analyzed by flow cytometry. Differentiation was performed resulting in osteogenic, chondrogenic and adipogenic cells. WJ-MSCs formed a homogenous monolayer of adherent spindle-shaped cells. Our results showed the high capacity of the proliferation of WJ-MSCs. Immunophenotyping further confirmed the purity of the isolated cells; their surface antigen expression showed the phenotypical properties like those of WJ-MSCs. The expanded cells were positive for CD 90, CD105, and CD44; they were negative for CD34 and HLA-DR surface markers. The cells had the adipocytic, osteocytic and chondrogenic differentiation capacity. The isolation and characterization of WJ-MSCs with high purity had been conducted, and the results were obtained in a short span. The present study has revealed the feasibility of the culture medium with high glucose and 15% FBS in isolation and proliferation of WJ-MSCs. When Wharton`s jelly pieces were put in the dry bottom of the flask, very effective separation of the MSCs was achieved.


Mesenchymal stem cells; Wharton`s jelly; Flow cytometry; Immunophenotyping

Full Text:



-El-Jawhari JJ, El-Sherbiny YM, Jones EA, McGonagle D. Mesenchymal stem cells,

Autoimmunity and rheumatoid arthritis. QJM. 2014; 107:505–14.

-Amorin B, Alegretti AP, Valim V, Pezzi A, Laureano AM, da Silva MA, et al. Mesenchymal stem cell therapy and acute graft-versus-host disease: a review. Hum Cell.

; 27:137–50.

- Wang D, Zhang H, Liang J, Li X, Feng X, Wang H, et al. Allogeneic mesenchymal stem cell transplantation in severe and refractory systemic lupus erythematosus: 4 years of experience. Cell Transplant. 2013; 22:2267–77.

- Kim MJ, Shin KS, Jeon JH, et al. Human chorionic- plate-derived mesenchymal stem cells and Whar-ton’s jelly-derived mesenchymal stem cells: a com-parative analyzis of their potential as placenta- derived stem cells. Cell Tissue Res 2011; 346:53-64.

- Taghizadeh RR, Cetrulo KJ, Cetrulo CL. Wharton’s Jelly stem cells: future clinical applications. Placenta 2011; 32 Suppl 4:S311-5.

-Witkowska-Zimny M, Wrobel E. Perinatal sources of mesenchymal stem cells: Wharton’s jelly, am-nion and chorion. Cell Mol Biol Lett 2011; 16:493- 514

- Abediankenari S, Ghasemi M, Kim YJ.Human leukocyte antigen-G expression on dendritic cells induced by transforming growth factor-beta1 and CD4+ T cells proliferation.Iran Biomed J. 2011; 15(1-2):1-5.

--Fong CY, Richards M, Manasi N, Biswas A, Bongso A. Comparative growth behaviour and characterization of stem cells from human Wharton’s jelly. Reprod Biomed Online 2007; 15: 708-718.

- Li G, Abediankenari S, Kim YJ, Campbell TB, Ito S, Graham-Evans B, Cooper S, Broxmeyer HE.TGF-beta combined with M-CSF and IL-4 induces generation of immune inhibitory cord blood dendritic cells capable of enhancing cytokine-induced ex vivo expansion of myeloid progenitors. Blood. 2007; 110(8):2872-9.

-Bunnell BA, Flaat M, Gagliardi C, Patel B, and Ripoll C. Adipose derived stem cells: Isolation, expansion and differentiation. Methods 2008; 45: 115-20.

-Wang H-S, Hung S-C, Peng S-T, et al. Mesenchymal stem cells in the Wharton's jelly of the human umbilical cord. Stem Cells 2004; 22: 1330-7.

-Sarugaser R, Lickorish D, Baksh D, Hosseini MM, Davies JE. Human umbilical cord perivascular (HUCPV) cells: a source of mesenchymal progenitors. Stem Cells 2005; 23: 220-9.

-Weiss ML, Medicetty S, Bledsoe AR, et al. Human umbilical cord matrix stem cells: preliminary characterization and effect of transplantation in a rodent model of Parkinson's disease. Stem Cells 2006; 24: 781-92.

-Lu L-L, Liu Y-J, Yang S-G, et al. Isolation and characterization of human umbilical cord mesenchymal stem cells with hematopoiesis supportive function and other potentials. Hematological 2006; 91: 1017-26.

-Karahuseyinoglu S, Cinar O, Kilic E, et al. Biology of stem cells in human umbilical cord stroma: in situ and in vitro surveys. Stem Cells 2007; 25: 319-31.

-Lund RD, Wang S, Lu B, et al. Cells isolated from umbilical cord tissue rescue photoreceptors and visual functions in a rodent model of retinal disease. Stem Cells 2007; 25: 602-11.

-Rocca GL, Anzalone R, Corrao S, et al. Isolation and characterization of Oct-4+/HLA-G+ mesenchymal stem cells from human umbilical cord matrix: differentiation potential and detection of new markers. Histochem Cell Biol 2009; 131: 267-82.

- Gnecchi M, He H, Liang OD, Melo LG, Morello F, Mu H, et al. Paracrine action accounts for marked protection of ischemic heart by Akt-modified mesenchymal stem cells. Nature Med. 2005; 11(4):367–8.

- Schuleri KH, Feigenbaum GS, Centola M, Weiss ES, Zimmet JM, Turney J, et al. Autologous mesenchymal stem cells produce reverse remodelling in chronic ischemic cardiomyopathy. Eur Heart J. 2009; 30(22):2722–32.

-Hatzistergos KE, Quevedo H, Oskouei BN, Hu Q, Feigenbaum GS, Margitich IS, et al. Bone marrow mesenchymal stem cells stimulate cardiac stem cell proliferation and differentiation. Circ Res. 2010; 107(7):913–22.

-Amado LC, Schuleri KH, Saliaris AP, Boyle AJ, Helm R, Oskouei B, et al. Multimodality noninvasive imaging demonstrates in vivo cardiac regeneration after mesenchymal stem cell therapy. J Am Coll Car diol. 2006; 48(10):2116–24.

-Karantalis V, DiFede DL, Gerstenblith G, Pham S, Symes J, Zambrano JP, et al. Autologous mesenchymal stem cells produce concordant improvements in regional function, tissue perfusion, and fibrotic burden when administered to patients undergoing coronary artery bypass grafting: the prospective randomized study of mesenchymal stem cell therapy in patients undergoing cardiac surgery (PROMETHEUS) trial. Circ Res. 2014; 114(8):1302–10.

- Heldman AW, DiFede DL, Fishman JE, Zambrano JP, Trachtenberg BH, Karantalis V, et al. Trans endocardial mesenchymal stem cells and mononuclear bone marrow cells for ischemic cardiomyopathy: the TAC-HFT randomized trial. JAMA. 2014; 311(1):62–73.

-Hare JM, Traverse JH, Henry TD, Dib N, Strumpf RK, Schulman SP, et al. A randomized, double-blind, placebo-controlled, dose-escalation study of intravenous adult human mesenchymal stem cells (prochymal) after acute myocardial infarction. J Am Coll car diol. 2009; 54(24):2277–86.

- Hare JM, DiFede DL, Castellanos AM, Florae V, Landin AM, El-Khorazaty J et al. Randomized Comparison of Allogeneic Vs. Autologous mesenchymal stem cells for Non-ischemic Dilated Cardiomyopathy: POSEIDON-DCM Trial. J Am Coll car diol. 2017; 69(5):526–37.

-Vater C, Kasten P, Stiehler M. Culture media for the differentiation of mesenchymal stromal cells. Acta Biomater 2011; 7(2): 463-77.

-Battula VL, Treml S, Abele H, Bühring HJ. Prospective isolation and characterization of mesenchymal stem cells from human placenta using a frizzled-9-specific monoclonal antibody. Differentiation 2008; 76(4): 326-36.

- Abediankenari S, Ghasemi M. Generation of immune inhibitory dendritic cells and CD4+T regulatory cells inducing by TGF-beta.Iran J Allergy Asthma Immunol. 2009 8(1):25-30.

- Sawangmake C,Osathanon T,Nowwarote N,P Pavasant, et al.Neurogenic differentiation of human dental pulp stem cells using different induction protocols.Oral Disease(2014)20,352-358

-Nekoei M, Azarpira N, Kamalifar S, et al. 2015.In vitro differentiation of human umbilical cord Wharton`s jelly mesenchymal stromal cells to insulin producing clusters,(3)7,640-648 .


  • There are currently no refbacks.

Creative Commons Attribution-NonCommercial 3.0

This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.