Seyed Noureddin Nematollahi-Mahani Corresponding Author
1- Department of Anatomy, Afzalipour Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran
2- Neuroscience Research Center, Kerman University of Medical Sciences, Kerman, Iran
Mohammad Rezazadehkermani
- Medical Students Research Center, Kerman University of Medical Sciences, Kerman, Iran
Mostafa Latifpour
1- Department of Anatomy, Afzalipour Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran
2- Neuroscience Research Center, Kerman University of Medical Sciences, Kerman, Iran
Parvin Salehinejad
1- Nursery School, Kerman University of Medical Sciences, Kerman, Iran
2- Institute of Bioscience, University Putra Malaysia, Kerman, Iran

Received: 12/30/2008 Accepted: 3/16/2009 - Publisher : Avicenna Research Institute

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Introduction: Stem-cell therapy has recently been proposed as a useful technique in the treatment of various illnesses, particularly degenerative diseases.Human Umbilical Cord Mesenchymal cells (hUCM) are among the stem cells which have received more attention in recent years.The current study was designed to investigate the culture conditions of these cells and to study some biological and biochemical properties of these cells, such as alkaline phosphatase activity, colony formation properties in hanging drops culture and the growth rate in various cell concentrations.Materials and Methods: Human umbilical cord was collected following a healthy cesarean section at the operation room of Afzalipour Educational Hospital. Matrix fragments were cultured by organ explant method. The attached cells at confluence of >80% were sub-cultured in new culture dishes and were seeded at a 1×106 density for morphologic evaluations. Upon colony formation of the cells, they were further stained to detect alkaline phosphatase enzyme activity. Furthermore, cells at a 1×105 density were cultured in hanging drops for 48 hours and subsequently alkaline phosphatase activity was evaluated in the resultant colonies. In addition, cells were seeded at various densities in 96-well culture plates and cell activity was measured by Wst-1 cell proliferation assay kit following 48 hours of culture incubation.Results: HUCM cells formed alkaline phosphatase positive colonies in culture, as well as in hanging drops. Cell activity was correlated with cell population at the start of cell cultivation. Increased concentration of cells at the beginning of culture led to increased cell activity upon 48 hours of incubation.Conclusion: HUCM cells expressed alkaline phosphatase enzyme in vitro and constituted colonies in hanging drops. In addition, hUCM cells showed higher activity when cultured in larger populations. It seems that hUCM cells resemble both embryonic and some adult stem cells. Therefore, further study on the characteristics of these cells could provide a basis for their application in regenerative medicine.

Keywords: Alkaline phosphatase, Cell proliferation, Cell therapy, Mesenchymal stem cells, Pluripotent stem cells, Umbilical cord

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Figures, Charts, Tables


  1. K?rbling M, Estrov Z. Adult stem cells for tissue repair - a new therapeutic concept? N Engl J Med. 2003;349 (6):570-82.   [PubMed]
  2. McElreavey KD, Irvine AI, Ennis KT, McLean WH. Isolation, culture and characterisation of fibroblast-like cells derived from the Whartons jelly portion of human umbilical cord. Biochem Soc Trans. 1991;19(1):29S.   [PubMed]
  3. Mitchell KE, Weiss ML, Mitchell BM, Martin P, Davis D, Morales L, et al. Matrix cells from Whartons jelly form neurons and glia. Stem Cells. 2003;21(1):50-60.   [PubMed]
  4. Kobayashi K, Kubota T, Aso T. Study on myofibro-blast differentiation in the stromal cells of Whartons jelly: expression and localization of alpha-smooth muscle actin. Early Hum Dev. 1998;51(3):223-33.   [PubMed]
  5. Kadner A, Hoerstrup SP, Tracy J, Breymann C, Maurus CF, Melnitchouk S, et al. Human umbilical cord cells: a new cell source for cardiovascular tissue engineering. Ann Thorac Surg. 2002;74(4):S1422-8.   [PubMed]
  6. Bailey MM, Wang L, Bode CJ, Mitchell KE, Detamore MS. A comparison of human umbilical cord matrix stem cells and temporomandibular joint condylar chondrocytes for tissue engineering temporoman-dibular joint condylar cartilage. Tissue Eng. 2007;13 (8):2003-10.   [PubMed]
  7. Medicetty S, Bledsoe AR, Fahrenholtz CB, Troyer D, Weiss ML. Transplantation of pig stem cells into rat brain: proliferation during the first 8 weeks. Exp Neurol. 2004;190(1):32-41.   [PubMed]
  8. Wang HS, Hung SC, Peng ST, Huang CC, Wei HM, Guo YJ, et al. Mesenchymal stem cells in the Whar-tons jelly of the human umbilical cord. Stem Cells. 2004;22(7):1330-7.   [PubMed]
  9. Weiss ML, Medicetty S, Bledsoe AR, Rachakatla RS, Choi M, Merchav S, et al. Human umbilical cord matrix stem cells: preliminary characterization and effect of transplantation in a rodent model of Parkin-sons disease. Stem Cells. 2006;24(3):781-92.   [PubMed]
  10. Carlin R, Davis D, Weiss M, Schultz B, Troyer D. Expression of early transcription factors Oct-4, Sox-2 and Nanog by porcine umbilical cord (PUC) matrix cells. Reprod Biol Endocrinol. 2006;4:8.   [PubMed]
  11. Jomura S, Uy M, Mitchell K, Dallasen R, Bode CJ, Xu Y. Potential treatment of cerebral global ischemia with Oct-4+ umbilical cord matrix cells. Stem Cells. 2007;25(1):98-106.   [PubMed]
  12. Hoynowski SM, Fry MM, Gardner BM, Leming MT, Tucker JR, Black L, et al. Characterization and differ-entiation of equine umbilical cord-derived matrix cells. Biochem Biophys Res Commun. 2007;362(2):347-53.   [PubMed]
  13. Tian X, Fu R, Deng L. [Method and conditions of isolation and proliferation of multipotent mesenchymal stem cells]. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2007;21(1):81-5. Chinese.   [PubMed]
  14. Troyer DL, Weiss ML. Whartons jelly-derived cells are a primitive stromal cell population. Stem Cells. 2008;26(3):591-9.   [PubMed]
  15. Cho PS, Messina DJ, Hirsh EL, Chi N, Goldman SN, Lo DP, et al. Immunogenicity of umbilical cord tissue derived cells. Blood. 2008;111(1):430-8.   [PubMed]
  16. Raio L, Cromi A, Ghezzi F, Passi A, Karousou E, Viola M, et al. Hyaluronan content of Whartons jelly in healthy and Down syndrome fetuses. Matrix Biol. 2005;24(2):166-74.   [PubMed]
  17. Sobolewski K, Ma?kowski A, Ba?kowski E, Jaworski S. Wharton s jelly as a reservoir of peptide growth factors. Placenta. 2005;26(10):747-52.   [PubMed]
  18. Karahuseyinoglu S, Cinar O, Kilic E, Kara F, Akay GG, Demiralp DO, et al. Biology of stem cells in human umbilical cord stroma: in situ and in vitro sur-veys. Stem Cells. 2007;25(2):319-31.   [PubMed]
  19. Yu X, Jin G, Yin X, Cho S, Jeon J, Lee S, et al. Isolation and characterization of embryonic stem-like cells derived from in vivo-produced cat blastocysts. Mol Reprod Dev. 2008;75(9):1426-32.   [PubMed]
  20. Guo Y, Mantel C, Hromas RA, Broxmeyer HE. Oct-4 is critical for survival/antiapoptosis of murine embry-onic stem cells subjected to stress: effects associated with Stat3/ survivin. Stem Cells. 2008;26(1):30-4.   [PubMed]
  21. Takechi K, Kuwabara Y, Mizuno M. Ultrastructural and immunohistochemical studies of Wharton s jelly umbilical cord cells. Placenta. 1993;14(2):235-45.   [PubMed]


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