@2024 Afarand., IRAN
ISSN: 2251-8215 Sarem Journal of Reproductive Medicine 2017;1(1):9-13
ISSN: 2251-8215 Sarem Journal of Reproductive Medicine 2017;1(1):9-13
Osteoblastic Differentiation of Amniotic Pluripotent Stem Cell
ARTICLE INFO
Article Type
Original ResearchAuthors
Mahmoodinia Maymand M. (1)Noruzinia M. (*)
(*) Medical Genetics Department, Medical Sciences, Tarbiat Modares University, Tehran, Iran
(1) Sarem Cell Research Center (SCRC), Sarem Women’s Hospital, Tehran, Iran
Correspondence
Article History
Received: September 27, 2015Accepted: January 12, 2016
ePublished: February 15, 2017
ABSTRACT
Aims
Amniotic fluid stem cells have lower ethical limitations than embryonic stem cells for the use in research and treatment. These cells show great self-renewal potential and can differentiate into the specialized cells of all three germ layers. The amniotic fluid stem cells display minimal risks of teratomas and very low immunogenicity. For these reasons, amniotic fluid appears as a promising alternative source for stem cell therapy. The objective of this study was to isolate the stem cells from amniotic fluid and differentiate them into the osteoblastic cells.
Materials & Methods An amniotic fluid sample (about 10ml) was collected from a healthy donor in Sarem women’s hospital (Tehran, Iran). After centrifugation, the cells were cultured in a DMEM medium supplemented with 20% FBS. The cell clones were observed after two weeks and were passaged to an osteoblastic differentiation medium. Alizarin red staining and RT-PCR for alkaline phosphatase and osteocalcin markers were used for confirmation of cellular differentiation.
Findings Stem cells were isolated from amniotic fluid. Phenotypically, these cells showed spindle-shaped morphology with a large nucleus. Following the induction of differentiation, they showed the expression of osteoblastic cells markers indicating their differentiation. The expression of those markers was confirmed by immunocytochemistry and RT-PCR.
Conclusion Amniotic stem cells have the ability to differentiate into the osteoblastic cells using an osteoblastic differentiation medium.
Materials & Methods An amniotic fluid sample (about 10ml) was collected from a healthy donor in Sarem women’s hospital (Tehran, Iran). After centrifugation, the cells were cultured in a DMEM medium supplemented with 20% FBS. The cell clones were observed after two weeks and were passaged to an osteoblastic differentiation medium. Alizarin red staining and RT-PCR for alkaline phosphatase and osteocalcin markers were used for confirmation of cellular differentiation.
Findings Stem cells were isolated from amniotic fluid. Phenotypically, these cells showed spindle-shaped morphology with a large nucleus. Following the induction of differentiation, they showed the expression of osteoblastic cells markers indicating their differentiation. The expression of those markers was confirmed by immunocytochemistry and RT-PCR.
Conclusion Amniotic stem cells have the ability to differentiate into the osteoblastic cells using an osteoblastic differentiation medium.
CITATION LINKS
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[15]Mao JJ, Giannobile WV, Helms JA, Hollister SJ, Krebsbach PH, Longaker MT, et al. Craniofacial tissue engineering by stem cells. J Dent Res. 2006;85(11):966-79.
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[17]Salasznyk RM, Williams WA, Boskey A, Batorsky A, Plopper GE. Adhesion to vitronectin and collagen I promotes osteogenic differentiation of human mesenchymal stem cells. J Biomed Biotechnol. 2004;2004(1):24-34.
[18]Liu XJ, Ren GH, Liao H, Yu L, Yuan L. Induced differentiation of adult human bone marrow derived mesenchymal stem cells in vitro toward osteoblasts. Di Yi Jun Yi Da Xue Xue Bao. 2004;24(4):408-11, 418.
[19]Tsai MT, Li WJ, Tuan RS, Chang WH. Modulation of osteogenesis in human mesenchymal stem cells by specific pulsed electromagnetic field stimulation. J Orthop Res. 2009;27(9):1169-74.
[20]Raouf A. Seth A. Discovery of osteoblast-associated genes using cDNA microarrays. Bone. 2002;30(3):463-71.
[2]Antonucci I, Iezzi I, Morizio E, Mastrangelo F, Pantalone A, Mattioli-Belmonte M, et al. Isolation of osteogenic progenitors from human amniotic fluid using a single step culture protocol. BMC Biotechnol, 2009;9:9.
[3]Cabral ACV, Ângelo PC, Leite HV, Pereira AK, Lopes APBM, Oliveira MB. Isolation, differentiation and biochemical aspects of amniotic fluid stem cell. Rev Assoc Med Bras. 2008;54(6):489-93.
[4]Gekas J, Walther G, Skuk D, Bujold E, Harvey I, Bertrand OF. In vitro and in vivo study of human amniotic fluid-derived stem cell differentiation into myogenic lineage. Clin Exp Med. 10(1):1-6.
[5]Zhang X, Chen X, Wang H, Liu S. Development of amniotic fluid-derived stem cell. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2008;22(7):864-8.
[6]Baksh D, Song L, Tuan RS. Adult mesenchymal stem cells: Characterization, differentiation, and application in cell and gene therapy. J Cell Mol Med. 2004;8(3):301-16.
[7]De Coppi P, Bartsch GJR, Siddiqui MM, Xu T, Santos CC, Perin L, et al. Isolation of amniotic stem cell lines with potential for therapy. Nat Biotechnol. 2007;25(1):100-6.
[8]Holden C. Versatile stem cells without the ethical baggage?. Science. 2007;315(5809):170.
[9]Kaviani A, Perry TE, Dzakovic A, Jennings RW, Ziegler MM, Fauza DO, Et al. The amniotic fluid as a source of cells for fetal tissue engineering. J Pediatr Surg. 2001;36(11):1662-5.
[10]Palumbo C, Ferretti M, Ardizzoni A, Zaffe D, Marotti G. Osteocyte-osteoclast morphological relationships and the putative role of osteocytes in bone remodeling. J Musculoskelet Neuronal Interact. 2001;1(4):327-32.
[11]Zuk PA. Tissue engineering craniofacial defects with adult stem cells? Are we ready yet?. Pediatr Res. 2008;63(5):478-86.
[12]Hee HT, Ismail HD, Lim CT, Goh JC, Wong HK. Effects of implantation of bone marrow mesenchymal stem cells, disc distraction and combined therapy on reversing degeneration of the intervertebral disc. J Bone Joint Surg Br. 2010;92(5):726-36.
[13]Thomas D, Kansara M. Epigenetic modifications in osteogenic differentiation and transformation. J Cell Biochem. 2006;98(4):757-69.
[14]Beloti, MM. Rosa AL. Osteoblast differentiation of human bone marrow cells under continuous and discontinuous treatment with dexamethasone. Braz Dent J. 2005;16(2):156-61.
[15]Mao JJ, Giannobile WV, Helms JA, Hollister SJ, Krebsbach PH, Longaker MT, et al. Craniofacial tissue engineering by stem cells. J Dent Res. 2006;85(11):966-79.
[16]Kassem M, Kristiansen M, Abdallah BM. Mesenchymal stem cells: Cell biology and potential use in therapy. Basic Clin Pharmacol Toxicol, 2004;95(5):209-14.
[17]Salasznyk RM, Williams WA, Boskey A, Batorsky A, Plopper GE. Adhesion to vitronectin and collagen I promotes osteogenic differentiation of human mesenchymal stem cells. J Biomed Biotechnol. 2004;2004(1):24-34.
[18]Liu XJ, Ren GH, Liao H, Yu L, Yuan L. Induced differentiation of adult human bone marrow derived mesenchymal stem cells in vitro toward osteoblasts. Di Yi Jun Yi Da Xue Xue Bao. 2004;24(4):408-11, 418.
[19]Tsai MT, Li WJ, Tuan RS, Chang WH. Modulation of osteogenesis in human mesenchymal stem cells by specific pulsed electromagnetic field stimulation. J Orthop Res. 2009;27(9):1169-74.
[20]Raouf A. Seth A. Discovery of osteoblast-associated genes using cDNA microarrays. Bone. 2002;30(3):463-71.