Research Article

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

Volume: 13 Number: 3 July 1, 2022
Acıbadem Üniversitesi Sağlık Bilimleri Dergisi Cover
Acıbadem Üniversitesi Sağlık Bilimleri Dergisi
PDF Download
EN

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

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.

Keywords

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

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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.
  8. 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

Details

Primary Language

English

Subjects

Biochemistry and Cell Biology (Other) , ​Internal Diseases , Pathology , Clinical Sciences (Other) , Obstetrics and Gynaecology

Journal Section

Research Article

Authors

Publication Date

July 1, 2022

Submission Date

January 6, 2022

Acceptance Date

January 22, 2022

Published in Issue

Year 2022 Volume: 13 Number: 3

DOI

https://doi.org/10.31067/acusaglik.1053582

IZ

https://izlik.org/JA57ME48XY

Cite

RIS / Bibtex
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.