MESENCHYMAL STROMAL CELLS FROM WHARTON’S JELLY EXPLANT CULTURE NOT ONLY CHARACTERIZES EMBRYONIC STEM CELLS BUT ALSO HAS GERM LAYER MARKERS
Main Article Content
Keywords
Explant, Germ layer markers, Mesenchymal stromal cells, Primitive nature, Wharton’s jelly
Abstract
Background and Objectives: Mesenchymal stromal cells (MSCs) have a unique property of immunomodulatory, homing and regenerative potential. It can also be isolated from several neonate tissues like placenta, umbilical cord and umbilical cord blood as they are considered primitive tissues. Human umbilical cord Wharton’s Jelly (hUCWJ) is a rich and promising source of MSCs due to its close association with embryonic tissue. It is a non-controversial and easily available source of human tissue. The aim of the study was to examine the expression of embryonic stem cells and germ layers markers in the neonate derived MSCs from hUCWJ.
Methods: The ex-plant isolated MSCs from hUCWJ were sub-cultured and the study was conducted at 3rd passage. After defining the isolated cells as MSCs (by morphology analysis, immunophenotyping by flowcytometer and tri-lineage differentiation), a semi-quantitative two-step reverse transcription polymerase chain reaction (RT-PCR) was performed to synthesize complementary DNA (cDNA). Finally using specific primers for embryonic stem cells and germ layer markers, PCR (Polymerase Chain Reaction) was performed on the cDNA and the products analyzed.
Results: A distinct expression of germ layer markers (nestin, TGFβ (transforming growth factor beta), GATA4) and embryonic stem cells markers (OCT4 (Octamer-binding transcription), NANOG (Nanog homeobox gene) and SOX2 (SRY (sex determining region Y)-box 2)) were observed on hUCWJ-MSCs
Interpretation & conclusions: A strong expression of endodermal marker could be observed compared to ectodermal and mesodermal markers. It also expresses embryonic stem cells markers supporting the pluripotency and primitive nature of the explant derived hUCWJ MSCs.
References
2. Bunnell BA, Flaat M, Gagliardi C, Patel B, Ripoll C. Adipose-derived stem cells: isolation, expansion and differentiation. Methods. 2008; 45: 115–120.
3. Ulrich D, Muralitharan R, Gargett CE. Toward the use of endometrial and menstrual blood mesenchymal stem cells for cell-based therapies. Expert OpinBiolTher. 2013; 13:1387–1400.
4. Roubelakis MG, Pappa KI, Bitsika V, Zagoura D, Vlahou A, Papadaki HA et al. Molecular and proteomic characterization of human mesenchymal stem cells derived from amniotic fluid: comparison to bone marrow mesenchymal stem cells. Stem Cells Dev. 2007; 16: 931–952.
5. Wang HS, Hung SC, Peng ST, Huang CC, Wei HM, Guo YJ, et al. Mesenchymal stem cells in the Wharton's jelly of the human umbilical cord. Stem Cells. 2004; 22:1330–1337.
6. Lee OK, Kuo TK, Chen WM, Lee KD, Hsieh SL, Chen TH. Isolation of multipotent mesenchymal stem cells from umbilical cord blood. Blood. 2004; 103:1669–1675.
7. Perry BC, Zhou D, Wu X, Yang FC, Byers MA, Chu TM et al. Collection, cryopreservation, and characterization of human dental pulp-derived mesenchymal stem cells for banking and clinical use. Tissue Eng Part C Methods. 2008; 14:149–56.
8. In 't Anker P, Scherjon S, Kleijburg-van der Keur C, de Groot-Swings G, Claas F, Fibbe W et al. Isolation of Mesenchymal Stem Cells of Fetal or Maternal Origin from Human Placenta. Stem Cells. 2004;22(7):1338-45.
9. Chamberlain G, Fox J, Ashton B, Middleton J. Concise Review: Mesenchymal Stem Cells: Their Phenotype, Differentiation Capacity, Immunological Features, and Potential for Homing. Stem Cells. 2007;25:2739–49
10. Marędziak M, Marycz K, Tomaszewski K, Kornicka K, Henry B. The Influence of Aging on the Regenerative Potential of Human Adipose Derived Mesenchymal Stem Cells. Stem Cells Internat. 2016;2016:1-15.
11. Troyer DL, Weiss ML.Wharton's jelly-derived cells are a primitive stromal cell population. Stem Cells. 2008;26:591-9
12. Boyer LA, Lee TI, Cole MF, Johnstone SE, Levine SS, ZuckerJPetal.Core transcriptional regulatory circuitry in human embryonic stem cells. Cell. 2005; 122:947-56.
13. Kim J, Jee M, Lee S, Han T, Kim B, Kang K et al. Regulation of Adipose Tissue Stromal Cells Behaviors by Endogenic Oct4 Expression Control. PLoS ONE. 2009;4(9):e7166.
14. Sharov A, Masui S, Sharova L, Piao Y, Aiba K, Matoba R et al. Identification of Pou5f1, Sox2, and Nanog downstream target genes with statistical confidence by applying a novel algorithm to time course microarray and genome-wide chromatin immunoprecipitation data. BMC Genomics. 2008;9(1):269.
15. Reim G, Brand M. Spiel-ohne-grenzen/pou2 mediates regional competence to respond to Fgf8 during zebrafish early neural development. Development. 2002;129:917–33.
16. Fang X, Yoon J, Li L, Yu W, Shao J, Hua D et al. The SOX2 response program in glioblastoma multiforme: an integrated ChIP-seq, expression microarray, and microRNA analysis. BMC Genom. 2011;12(1).
17. Sun A, Liu CJ, Sun ZQ, Wei Z. NANOG: A promising target for digestive malignant tumors. World J Gastroenterol. 2014;20(36):13071.
18. Riekstina U, Cakstina I, Parfejevs V, Hoogduijn M, Jankovskis G, Muiznieks I et al. Embryonic Stem Cell Marker Expression Pattern in Human Mesenchymal Stem Cells Derived from Bone Marrow, Adipose Tissue, Heart and Dermis. Stem Cell Rev. 2009;5(4):378-386.
19. Lengner C, Welstead G, Jaenisch R. The pluripotency regulator Oct4: A role in somatic stem cells?.Cell Cycle. 2008;7(6):725-8.
20. Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006;8(4):315-7.
21. Caplan AI. Mesenchymal Stem Cells: Time to Change the Name. Stem Cells Transl Med. 2017;6(6):1445-51.
22. Karahuseyinoglu S, Cinar O, Kilic E, Kara F, Akay G, Demiralp D et al. Biology of Stem Cells in Human Umbilical Cord Stroma: In Situ and In Vitro Surveys. Stem Cells. 2007;25(2):319-31.
23. Marcus A, Woodbury D. Fetal stem cells from extra-embryonic tissues: do not discard. J Cell Mol Med. 2008;12(3):730-42.
24. Hendijani F. Explant culture: An advantageous method for isolation of mesenchymal stem cells from human tissues. Cell Prolif. 2017; 50(2): e12334.
25. Limana F, Zacheo A, Mocini D, Mangoni A, Borsellino G, Diamantini A et al. Identification of Myocardial and Vascular Precursor Cells in Human and Mouse Epicardium. Circ Res. 2007;101(12):1255-65.
26. Bhartiya D. Are Mesenchymal cells indeed pluripotent stem cells or just stromal cells? OCT-4 and VSELs biology has led to better understanding. Stem Cells Int. 2013; 2013:547501.
27. Arnold K, Sarkar A, Yram M, Polo J, Bronson R, Sengupta S et al. Sox2+ Adult Stem and Progenitor Cells Are Important for Tissue Regeneration and Survival of Mice. Cell Stem Cell. 2011;9(4):317-29.
28. Cao D. The origin of cancer stem cells. Frontiers in Bioscience. 2012;S4(3):819-30.
29. Malik B. Cancer stems cells and resistance to chemo and radio therapy. Front in Bioscien. 2012;E4(1):2142.
30. Pierantozzi E, Gava B, Manini I, Roviello F, Marotta G, Chiavarelli M et al. Pluripotency Regulators in Human Mesenchymal Stem Cells: Expression of NANOG But Not of OCT-4 and SOX-2. Stem Cells Dev. 2011;20(5):915-23.
Article Sidebar
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
All articles published in JPTCP are licensed under Copyright Creative Commons Attribution-NonCommercial 4.0 International License.
T Paras Singh
Associate Professor, Department of Biochemistry, Shija Academy of Health Sciences, Imphal West, Manipur, India
M L Sherpa
Professor, Department of Biochemistry, Sikkim Manipal Institute of Medical Sciences, Sikkim Manipal University, Gangtok, Sikkim, India
Anup Pradhan
Professor, Department of Obstetrics and Gynecology, Sikkim Manipal Institute of Medical Sciences, Sikkim Manipal University, Gangtok, Sikkim, India
T A Singh
Professor, Department of Biochemistry, Shija Academy of Health Sciences, Imphal West, Manipur, India