Comparative Characterization of Cultured Human Term Amnion Epithelial and Mesenchymal Stromal Cells for Application in Cell Therapy
| Autor: | Roland Zimmermann, Ajit S. Mallik, Steffen M. Zeisberger, Grozdana Bilic, Andreas H. Zisch |
|---|---|
| Přispěvatelé: | University of Zurich, Bilic, G |
| Jazyk: | angličtina |
| Rok vydání: | 2008 |
| Předmět: |
Pathology
medicine.medical_specialty 2747 Transplantation Cell Survival Biomedical Engineering Cell Culture Techniques Cell- and Tissue-Based Therapy 2204 Biomedical Engineering lcsh:Medicine 610 Medicine & health Stem cell factor Biology Mesenchymal Stem Cell Transplantation 1307 Cell Biology Andrology medicine Humans CD90 Telomerase reverse transcriptase Amnion RNA Messenger Progenitor cell 10026 Clinic for Obstetrics Cell Shape Cells Cultured Cell Proliferation Transplantation Stem Cell Factor Reverse Transcriptase Polymerase Chain Reaction Mesenchymal stem cell lcsh:R Cell Differentiation Epithelial Cells Mesenchymal Stem Cells Cell Biology Embryonic stem cell medicine.anatomical_structure 10076 Center for Integrative Human Physiology Antigens Surface embryonic structures cardiovascular system 570 Life sciences biology Stromal Cells Octamer Transcription Factor-3 Biomarkers |
| Zdroj: | Cell Transplantation, Vol 17 (2008) |
| ISSN: | 1555-3892 0963-6897 |
| Popis: | Emerging evidence suggests human amnion tissue as a valuable source of two distinct types of pluripotent cells, amnion epithelial cells (hAECs) and mesenchymal stromal cells (hAMSCs), for applications in cell replacement therapy. For some approaches, it may be necessary to culture and differentiate these cells before they can be transplanted. No systematic attempt has been yet made to determine the quantity and quality of amnion cells after isolation and culture. We looked at amnion cell isolates from 27 term placentas. Following our optimized protocol, primary yields were 6.3 × 106 hAECs and 1.7 × 106 hAMSCs per gram amnion. All 27 cases gave vital cultures of hAMSCs, while one third of cases (9 of 27) failed to give adherent cultures of hAECs. Primary cultures contained significantly more proliferating than apoptotic cells (hAECs: 16.4% vs. 4.0%; hAMSCs: 9.5% vs. 2.4%). Neither hAECs nor hAMSCs were clonogenic. They showed slow proliferation that almost stopped beyond passage 5. Microscopic follow-up revealed that hAEC morphology gradually changed towards mesenchymal phenotype over several passages. Flow cytometric characterization of primary cultures showed expression of mesenchymal progenitor markers CD73, CD90, CD105, and CD166, as well as the embryonic stem cell markers SSEA-3 and -4 on both amnion cell types. These profiles were grossly maintained in secondary cultures. Reverse transcriptase-PCR analysis exhibited transcripts of Oct-3/4 and stem cell factor in primary and secondary cultures of all cases, but no telomerase reverse transcriptase. Immunocytochemistry confirmed translation into Oct-3/4 protein in part of hAEC cultures, but not in hAMSCs. Further, both amnion cell types stained for CD90 and SSEA-4. Osteogenic induction studies with amnion cells from four cases showed significantly stronger differentiation of hAECs than hAMSCs; this capacity to differentiate greatly varied between cases. In conclusion, hAECs and hAMSCs in culture exhibit and maintain a similar marker profile of mesenchymal progenitors. hAECs were found as a less reliable source than hAMSCs and altered morphology during subculture. |
| Databáze: | OpenAIRE |
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