A Comparative Study of Growth Kinetics, In Vitro Differentiation Potential and Molecular Characterization of Fetal Adnexa Derived Caprine Mesenchymal Stem Cells

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2016 Jun 4

PMID 27257959

Citations 10

Authors

Anjali Somal

Irfan A Bhat

Indu B

Sriti Pandey

Bibhudatta S K Panda

Nipuna Thakur

Mihir Sarkar

Vikash Chandra

G Saikumar

G Taru Sharma

Affiliations

Soon will be listed here.

Abstract

The present study was conducted with an objective of isolation, in vitro expansion, growth kinetics, molecular characterization and in vitro differentiation of fetal adnexa derived caprine mesenchymal stem cells. Mid-gestation gravid caprine uteri (2-3 months) were collected from abattoir to derive mesenchymal stem cells (MSCs) from fetal adnexa {amniotic fluid (cAF), amniotic sac (cAS), Wharton's jelly (cWJ) and cord blood (cCB)} and expanded in vitro. These cultured MSCs were used at the 3rd passage (P3) to study growth kinetics, localization as well as molecular expression of specific surface antigens, pluripotency markers and mesenchymal tri-lineage differentiation. In comparison to cAF and cAS MSCs, cWJ and cCB MSCs showed significantly (P<0.05) higher clonogenic potency, faster growth rate and low population doubling (PDT) time. All the four types of MSCs were positive for alkaline phosphatase (AP) and differentiated into chondrogenic, osteogenic, and adipogenic lineages. These stem cells expressed MSC surface antigens (CD73, CD90 and CD105) and pluripotency markers (Oct4, Sox2, Nanog, KLF, cMyc, FoxD3) but did not express CD34, a hematopoietic stem cell marker (HSC) as confirmed by RT-PCR, immunocytochemistry and flow cytometric analysis. The relative mRNA expression of MSC surface antigens (CD73, CD90 and CD105) was significantly (P<0.05) higher in cWJ MSCs compared to the other cell lines. The mRNA expression of Oct4 was significantly (P<0.05) higher in cWJ, whereas mRNA expression of KLF and cMyc was significantly (P<0.05) higher in cWJ and cAF than that of cAS and cCB. The comparative assessment revealed that cWJ MSCs outperformed MSCs from other sources of fetal adnexa in terms of growth kinetics, relative mRNA expression of surface antigens, pluripotency markers and tri-lineage differentiation potential, hence, these MSCs could be used as a preferred source for regenerative medicine.

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References

Reynolds L, Borowicz P, Vonnahme K, Johnson M, Grazul-Bilska A, Wallace J . Animal models of placental angiogenesis. Placenta. 2005; 26(10):689-708. DOI: 10.1016/j.placenta.2004.11.010. View

Mann A, Yadav R, Singh J, Kumar D, Singh B, Yadav P . Culture, characterization and differentiation of cells from buffalo (Bubalus bubalis) amnion. Cytotechnology. 2012; 65(1):23-30. PMC: 3536876. DOI: 10.1007/s10616-012-9464-z. View

Udehiya R, Amarpal , Aithal H, Kinjavdekar P, Pawde A, Singh R . Comparison of autogenic and allogenic bone marrow derived mesenchymal stem cells for repair of segmental bone defects in rabbits. Res Vet Sci. 2013; 94(3):743-52. DOI: 10.1016/j.rvsc.2013.01.011. View

Kern S, Eichler H, Stoeve J, Kluter H, Bieback K . Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells. 2006; 24(5):1294-301. DOI: 10.1634/stemcells.2005-0342. View

Roubelakis M, Trohatou O, Anagnou N . Amniotic fluid and amniotic membrane stem cells: marker discovery. Stem Cells Int. 2012; 2012:107836. PMC: 3372280. DOI: 10.1155/2012/107836. View

Iacono E, Cunto M, Zambelli D, Ricci F, Tazzari P, Merlo B . Could fetal fluid and membranes be an alternative source for mesenchymal stem cells (MSCs) in the feline species? A preliminary study. Vet Res Commun. 2012; 36(2):107-18. DOI: 10.1007/s11259-012-9520-3. View

Hoynowski S, Fry M, Gardner B, Leming M, Tucker J, Black L . Characterization and differentiation of equine umbilical cord-derived matrix cells. Biochem Biophys Res Commun. 2007; 362(2):347-53. DOI: 10.1016/j.bbrc.2007.07.182. View

Garcia-Castro I, Garcia-Lopez G, Avila-Gonzalez D, Flores-Herrera H, Molina-Hernandez A, Portillo W . Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons. PLoS One. 2016; 10(12):e0146082. PMC: 4697857. DOI: 10.1371/journal.pone.0146082. View

Ilancheran S, Moodley Y, Manuelpillai U . Human fetal membranes: a source of stem cells for tissue regeneration and repair?. Placenta. 2008; 30(1):2-10. DOI: 10.1016/j.placenta.2008.09.009. View

10.

Gao Y, Zhu Z, Zhao Y, Hua J, Ma Y, Guan W . Multilineage potential research of bovine amniotic fluid mesenchymal stem cells. Int J Mol Sci. 2014; 15(3):3698-710. PMC: 3975362. DOI: 10.3390/ijms15033698. View

11.

Whealands Smith R, Werling N, Dakin S, Alam R, Goodship A, Dudhia J . Beneficial effects of autologous bone marrow-derived mesenchymal stem cells in naturally occurring tendinopathy. PLoS One. 2013; 8(9):e75697. PMC: 3783421. DOI: 10.1371/journal.pone.0075697. View

12.

Sarugaser R, Lickorish D, Baksh D, Hosseini M, Davies J . Human umbilical cord perivascular (HUCPV) cells: a source of mesenchymal progenitors. Stem Cells. 2005; 23(2):220-9. DOI: 10.1634/stemcells.2004-0166. View

13.

Duff S, Li C, Garland J, Kumar S . CD105 is important for angiogenesis: evidence and potential applications. FASEB J. 2003; 17(9):984-92. DOI: 10.1096/fj.02-0634rev. View

14.

Airas L, Hellman J, Salmi M, Bono P, Puurunen T, Smith D . CD73 is involved in lymphocyte binding to the endothelium: characterization of lymphocyte-vascular adhesion protein 2 identifies it as CD73. J Exp Med. 1995; 182(5):1603-8. PMC: 2192217. DOI: 10.1084/jem.182.5.1603. View

15.

Lange-Consiglio A, Corradetti B, Bizzaro D, Magatti M, Ressel L, Tassan S . Characterization and potential applications of progenitor-like cells isolated from horse amniotic membrane. J Tissue Eng Regen Med. 2011; 6(8):622-35. DOI: 10.1002/term.465. View

16.

Filioli Uranio M, Valentini L, Lange-Consiglio A, Caira M, Guaricci A, LAbbate A . Isolation, proliferation, cytogenetic, and molecular characterization and in vitro differentiation potency of canine stem cells from foetal adnexa: a comparative study of amniotic fluid, amnion, and umbilical cord matrix. Mol Reprod Dev. 2011; 78(5):361-73. DOI: 10.1002/mrd.21311. View

17.

Sartore S, Lenzi M, Angelini A, Chiavegato A, Gasparotto L, De Coppi P . Amniotic mesenchymal cells autotransplanted in a porcine model of cardiac ischemia do not differentiate to cardiogenic phenotypes. Eur J Cardiothorac Surg. 2005; 28(5):677-84. DOI: 10.1016/j.ejcts.2005.07.019. View

18.

Rossi B, Merlo B, Colleoni S, Iacono E, Tazzari P, Ricci F . Isolation and in vitro characterization of bovine amniotic fluid derived stem cells at different trimesters of pregnancy. Stem Cell Rev Rep. 2014; 10(5):712-24. DOI: 10.1007/s12015-014-9525-0. View

19.

Toda A, Okabe M, Yoshida T, Nikaido T . The potential of amniotic membrane/amnion-derived cells for regeneration of various tissues. J Pharmacol Sci. 2007; 105(3):215-28. DOI: 10.1254/jphs.cr0070034. View

20.

Mohanty N, Gulati B, Kumar R, Gera S, Kumar P, Somasundaram R . Immunophenotypic characterization and tenogenic differentiation of mesenchymal stromal cells isolated from equine umbilical cord blood. In Vitro Cell Dev Biol Anim. 2014; 50(6):538-48. DOI: 10.1007/s11626-013-9729-7. View