Research Article
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The Assessment of Mesenchymal Stem Cells Characteristics in Cultured Amniotic Fluid Cells

Year 2022, , 299 - 308, 01.07.2022
https://doi.org/10.31067/acusaglik.1053582

Abstract

OBJECTİVES: To evaluate the morphological characteristics, immunophenotypic features, osteogenic, adipogenic, and chondrogenic differentiation capacities of in-vitro cultured amniotic cells during the culture process based on mesenchymal stem cell (MSC).
MATERİAL/METHOD: This study used waste cells remaining after fetal karyotype determination with second-trimester amniotic cell culture. Cultured cells were morphologically evaluated daily. Selective subculture (SSC) was produced by the scraping method. Immunophenotypic features of primary and subculture amniotic cells were evaluated based on specific surface antigens. Specific growth mediums were used to assess their osteogenic, adipogenic, and chondrogenic differentiation abilities. Differentiation was confirmed using specific staining.
RESULTS: Fifteen cases with 46,XY fetal cytogenetic analyses at 16+3-19+5 gestational weeks were included in the study. Amniotic fluid cells were evaluated morphologically in three classes as Amniotic fluid specific cell-type(AF), Fibroblastoid cell-type(FB1, FB2), and Epithelioid cell-type(E). The first cell adhesions occurred in the first 24-48 hours(36%) of cell culture, while the first colonies within three days(82%). SSC was produced in AF-SSC(n=4) and FB2-SSC(n=6) directions by mechanical scraping. Primary amniotic fluid cells and AF/FB-SSCs were positive for MSC immunophenotypic markers CD29,CD73,CD166,CD44,CD49e,CD90, while negative for CD34,CD45, and HLA-DR. Cells were shown to have osteogenic (Alizarin Red-S) and chondrogenic (Alcian Blue) differentiation potentials based on histochemical staining, while adipogenic (Oil Red-O) differentiation was not obtained.
CONCLUSION: We believe that preliminary protocols and experiences to obtain MSCs from amniotic fluid-derived cells produced under routine prenatal diagnosis cell culture conditions can enable many promising pre-clinical/clinical studies and be the pioneer of "stem cell therapy applications" in many "incurable" diseases in the clinic.

Supporting Institution

OMÜ rektörlük projeleri

Project Number

T-393/394

Thanks

Acknowledgements: The author wishes to thank thesis advisor, Prof. Dr. Gonul OGUR for guiding.

References

  • Fortier LA. Stem cells: classifications, controversies, and clinical applications. Vet Surg 2005;34(5):415-23. doi:10.1111/j.1532-950X.2005.00063.x
  • De Coppi P, Bartsch G Jr, Siddiqui MM, et al. Isolation of amniotic stem cell lines with potential for therapy. Nat Biotechnol 2007;25(1):100-6. doi:10.1038/nbt1274
  • In 't Anker PS, Scherjon SA, Kleijburg-van der Keur C, et al. Amniotic fluid as a novel source of mesenchymal stem cells for therapeutic transplantation. Blood 2003;102(4):1548-9. doi:10.1182/blood-2003-04-1291
  • Kaviani A, Guleserian K, Perry TE, et al. Fetal tissue engineering from amniotic fluid. J Am Coll Surg 2003;196(4):592-7. doi:10.1016/s1072-7515(02)01834-3
  • Kaviani A, Perry TE, Dzakovic A, et al. The amniotic fluid as a source of cells for fetal tissue engineering. Journal of Pediatric Surgery 2001; 36:1662–1665. doi:10.1053/jpsu.2001.27945
  • Pittenger MF, Mackay AM, Beck SC, et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999;284:143–147. doi:10.1126/science.284.5411.143
  • In 't Anker PS, Noort WA, Scherjon SA, Kleijburg-van der Keur C, Kruisselbrink AB, van Bezooijen RL, et al. Mesenchymal stem cells in human second-trimester bone marrow, liver, lung, and spleen exhibit a similar immunophenotype but a heterogeneous multilineage differentiation potential. Haematologica 2003;88(8):845-52.
  • Portmann-Lanz CB, Schoeberlein A, Huber A, et al. Placental mesenchymal stem cells as potential autologous graft for pre- and perinatal neuroregeneration. Am J Obstet Gynecol 2006;194(3):664-73. doi:10.1016/j.ajog.2006.01.101
  • In 't Anker PS, Scherjon SA, Kleijburg-van der Keur C, et al. Isolation of mesenchymal stem cells of fetal or maternal origin from human placenta. Stem Cells 2004;22(7):1338-45. doi:10.1634/stemcells.2004-0058
  • Tsai MS, Hwang SM, Tsai YL, et al. Clonal amniotic fluid-derived stem cells Express characteristics of both mesenchymal and neural stem cells. Biol. Reprod 2006;74;545–551. doi:10.1095/biolreprod.105.046029
  • Bossolasco P, Montemurro T, Cova L, et al. Molecular and phenotypic characterization of human amniotic fluid cells and their differentiation potential. Cell Res 2006;16(4):329-36. doi:10.1038/sj.cr.7310043
  • Delo DM, De Coppi P, Bartsch G Jr, et al. Amniotic fluid and placental stem cells. Methods Enzymol 2006;419:426-38. doi:10.1016/S0076-6879(06)19017-5
  • Tsai MS, Lee JL, Chang YJ, et al. Isolation of human multipotent mesenchymal stem cells from second trimester amniotic fluid using a novel two-stage culture protocol. Hum. Reprod 2004;19:1450–1457. doi:10.1093/humrep/deh279
  • Lev R, Spicer SS. Specific staining of sulfate groups with Alcian blue at low pH. J. Histochem. Cytochem 1964;12:309. doi:10.1177/12.4.309
  • Kim J, Lee Y, Kim H, et al. Human amniotic fluid-derived stem cells have characteristics of multipotent stem cells. Cell Prolif 2007;40:75–90. doi:10.1111/j.1365-2184.2007.00414.x
  • http://www.protocol-online.org/prot/Histology/Staining/index.html
  • Gosden CM. Amniotic fluid cell types and culture. Br Med Bull 1983;39:348-54. doi:10.1093/oxfordjournals.bmb.a071847
  • Phillips CN, McCue PA, Whitsett CF, et al. Differentiation in human chorionic villus cultures: hCG and HLA expression. Prenat Diagn 1989;9(4):227-42. doi:10.1002/pd.1970090403
  • Hoehn H, Bryant EM, Karp LE, et al. Cultivated cells from diagnostic amniocentesis in second trimester pregnancies. I. Clonal morphology and growth potential. Pediat Res 1974;8:746-754. doi:10.1203/00006450-197408000-00003
  • Deedwania P, Deka D, Mohanty S, et al. Isolation and characterization of mesenchymal stem cells derived from amniotic fluid: A prospective study. Int J Mol Immuno Oncol 2020;5(2):67-72. doi:10.25259/IJMIO_22_2019
  • Prusa AR, Hengstschläger M. Amniotic fluid cells and human stem cell research: a new connection. Med Sci Monit 2002;8:253–257.
  • Polgar K, Abel G, Laczko J, et al. Immunocytochemical characterization of amniotic fluid macrophages in cases of fetal neural tube defects. Am J Clin Pathol 1987;87(1):37-42. doi:10.1093/ajcp/87.1.37
  • Kunisaki SM, Armant M, Kao GS, et al. Tissue engineering from human mesencymal amnocytes: a prelude to clinical trials. J Pediatr Surg. 2007;42:974-980. doi:10.1016/j.jpedsurg.2007.01.031
  • Miki T, Lehmann T, Cai H, et al. Stem cell characteristics of amniotic epithelial cells. Stem Cells 2005;23(10):1549-59. doi:10.1634/stemcells.2004-0357
  • Ichinose S, Tagami M, Muneta T, et al. Morphological examination during in vitro cartilage formation by human mesenchymal stem cells. Cell Tissue Res 2005;322(2):217-26. doi:10.1007/s00441-005-1140-6
Year 2022, , 299 - 308, 01.07.2022
https://doi.org/10.31067/acusaglik.1053582

Abstract

Project Number

T-393/394

References

  • Fortier LA. Stem cells: classifications, controversies, and clinical applications. Vet Surg 2005;34(5):415-23. doi:10.1111/j.1532-950X.2005.00063.x
  • De Coppi P, Bartsch G Jr, Siddiqui MM, et al. Isolation of amniotic stem cell lines with potential for therapy. Nat Biotechnol 2007;25(1):100-6. doi:10.1038/nbt1274
  • In 't Anker PS, Scherjon SA, Kleijburg-van der Keur C, et al. Amniotic fluid as a novel source of mesenchymal stem cells for therapeutic transplantation. Blood 2003;102(4):1548-9. doi:10.1182/blood-2003-04-1291
  • Kaviani A, Guleserian K, Perry TE, et al. Fetal tissue engineering from amniotic fluid. J Am Coll Surg 2003;196(4):592-7. doi:10.1016/s1072-7515(02)01834-3
  • Kaviani A, Perry TE, Dzakovic A, et al. The amniotic fluid as a source of cells for fetal tissue engineering. Journal of Pediatric Surgery 2001; 36:1662–1665. doi:10.1053/jpsu.2001.27945
  • Pittenger MF, Mackay AM, Beck SC, et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999;284:143–147. doi:10.1126/science.284.5411.143
  • In 't Anker PS, Noort WA, Scherjon SA, Kleijburg-van der Keur C, Kruisselbrink AB, van Bezooijen RL, et al. Mesenchymal stem cells in human second-trimester bone marrow, liver, lung, and spleen exhibit a similar immunophenotype but a heterogeneous multilineage differentiation potential. Haematologica 2003;88(8):845-52.
  • Portmann-Lanz CB, Schoeberlein A, Huber A, et al. Placental mesenchymal stem cells as potential autologous graft for pre- and perinatal neuroregeneration. Am J Obstet Gynecol 2006;194(3):664-73. doi:10.1016/j.ajog.2006.01.101
  • In 't Anker PS, Scherjon SA, Kleijburg-van der Keur C, et al. Isolation of mesenchymal stem cells of fetal or maternal origin from human placenta. Stem Cells 2004;22(7):1338-45. doi:10.1634/stemcells.2004-0058
  • Tsai MS, Hwang SM, Tsai YL, et al. Clonal amniotic fluid-derived stem cells Express characteristics of both mesenchymal and neural stem cells. Biol. Reprod 2006;74;545–551. doi:10.1095/biolreprod.105.046029
  • Bossolasco P, Montemurro T, Cova L, et al. Molecular and phenotypic characterization of human amniotic fluid cells and their differentiation potential. Cell Res 2006;16(4):329-36. doi:10.1038/sj.cr.7310043
  • Delo DM, De Coppi P, Bartsch G Jr, et al. Amniotic fluid and placental stem cells. Methods Enzymol 2006;419:426-38. doi:10.1016/S0076-6879(06)19017-5
  • Tsai MS, Lee JL, Chang YJ, et al. Isolation of human multipotent mesenchymal stem cells from second trimester amniotic fluid using a novel two-stage culture protocol. Hum. Reprod 2004;19:1450–1457. doi:10.1093/humrep/deh279
  • Lev R, Spicer SS. Specific staining of sulfate groups with Alcian blue at low pH. J. Histochem. Cytochem 1964;12:309. doi:10.1177/12.4.309
  • Kim J, Lee Y, Kim H, et al. Human amniotic fluid-derived stem cells have characteristics of multipotent stem cells. Cell Prolif 2007;40:75–90. doi:10.1111/j.1365-2184.2007.00414.x
  • http://www.protocol-online.org/prot/Histology/Staining/index.html
  • Gosden CM. Amniotic fluid cell types and culture. Br Med Bull 1983;39:348-54. doi:10.1093/oxfordjournals.bmb.a071847
  • Phillips CN, McCue PA, Whitsett CF, et al. Differentiation in human chorionic villus cultures: hCG and HLA expression. Prenat Diagn 1989;9(4):227-42. doi:10.1002/pd.1970090403
  • Hoehn H, Bryant EM, Karp LE, et al. Cultivated cells from diagnostic amniocentesis in second trimester pregnancies. I. Clonal morphology and growth potential. Pediat Res 1974;8:746-754. doi:10.1203/00006450-197408000-00003
  • Deedwania P, Deka D, Mohanty S, et al. Isolation and characterization of mesenchymal stem cells derived from amniotic fluid: A prospective study. Int J Mol Immuno Oncol 2020;5(2):67-72. doi:10.25259/IJMIO_22_2019
  • Prusa AR, Hengstschläger M. Amniotic fluid cells and human stem cell research: a new connection. Med Sci Monit 2002;8:253–257.
  • Polgar K, Abel G, Laczko J, et al. Immunocytochemical characterization of amniotic fluid macrophages in cases of fetal neural tube defects. Am J Clin Pathol 1987;87(1):37-42. doi:10.1093/ajcp/87.1.37
  • Kunisaki SM, Armant M, Kao GS, et al. Tissue engineering from human mesencymal amnocytes: a prelude to clinical trials. J Pediatr Surg. 2007;42:974-980. doi:10.1016/j.jpedsurg.2007.01.031
  • Miki T, Lehmann T, Cai H, et al. Stem cell characteristics of amniotic epithelial cells. Stem Cells 2005;23(10):1549-59. doi:10.1634/stemcells.2004-0357
  • Ichinose S, Tagami M, Muneta T, et al. Morphological examination during in vitro cartilage formation by human mesenchymal stem cells. Cell Tissue Res 2005;322(2):217-26. doi:10.1007/s00441-005-1140-6
There are 25 citations in total.

Details

Primary Language English
Subjects Biochemistry and Cell Biology (Other), ​Internal Diseases, Pathology, Clinical Sciences (Other), Obstetrics and Gynaecology
Journal Section Research Article
Authors

Ozlem Sezer 0000-0001-5727-7965

Gönül Oğur 0000-0002-9944-4423

Project Number T-393/394
Publication Date July 1, 2022
Submission Date January 6, 2022
Published in Issue Year 2022

Cite

EndNote Sezer O, Oğur G (July 1, 2022) The Assessment of Mesenchymal Stem Cells Characteristics in Cultured Amniotic Fluid Cells. Acıbadem Üniversitesi Sağlık Bilimleri Dergisi 13 3 299–308.