2011 - CTS-IXA


This page contains exclusive content for the member of the following sections: TTS, CTS, IXA. Log in to view.

Parallel Session 2- Stem Cells (Cell Track)

4.117 - Generation of induced pluripotent stem cells from Amnion Epithelial cells

Presenter: Roberto, Gramignoli, Pittsburgh, United States
Authors: Roberto Gramignoli1, Marc C. Hansel1, Fabio Marongiu2, William L. Blake3, Alejandro Soto-Gutierrez5, Kenneth Dorko1, Julio C. Davila4, Stephen C. Strom1

117

Generation of induced pluripotent stem cells from Amnion Epithelial cells

Roberto Gramignoli1, Marc C. Hansel1, Fabio Marongiu2, William L. Blake3, Alejandro Soto-Gutierrez5, Kenneth Dorko1, Julio C. Davila4, Stephen C. Strom1

1Pathology, University of Pittsburgh, Pittsburgh, PA, United States; 2Biomedical Sciences and Technologies, Università degli Studi di Cagliari, Cagliari, Italy; 3Genetically Modified Models Center of Emphasis, Pfizer, Groton, CT; 4Pfizer, St. Louis, MO; 5Surgery, University of Pittsburgh Medical Center, Pittsburgh, PA, United States

Placenta is a readily available and non-controversial source of cells that could be used in regenerative medicine. Human Amniotic Epithelial (AE) cells and induced pluripotent stem (iPS) cell technologies would seem to be devoid of most if not all of the ethical concerns associated with ES cells. We generated of iPS cell lines from primary human AE (AE-iPS). After exposure to lentiviral constructs carrying the reprogramming factors, cells with appropriate morphology were collected and were analyzed for markers of pluripotency including Oct4, Nanog, Sox2 and alkaline phosphatase. A profile of AE was conducted pre- and post-reprogramming by Flow Cytometry for surface markers including: SSEA-3, SSEA-4, TRA1-60, TRA1-81, HLA-ABC, CD24, CD29, CD34, CD44, CD49f, CD73, CD105, CD90, CD117, CD133/2, CD146, CD166, EpCAM and ABCG2. FACS analysis revealed a morphological difference in forward scatter characterized by a compartment with a higher side scatter in AE, not present in AE-iPS thought to indicate a different level of cellular complexity and granularity. The percentage of SSEA-3, SSEA-4, TRA1-60 and TRA1-81 positive cells after reprogramming was dramatically increased in AE-iPS cells (30-90%). We also observed that stem cell marker such as CD133/2 was highly expressed only after the reprogramming procedure. Colonies generated have morphological features and the surface marker and gene expression profile of fully reprogrammed cells and also form teratomas upon transplantation. These studies confirm that iPS lines can be generated from primary human amnion. Protocols to differentiate these cells as previously published by our group on AE cells toward a hepatic phenotype are in progress.


Important Disclaimer

By viewing the material on this site you understand and accept that:

  1. The opinions and statements expressed on this site reflect the views of the author or authors and do not necessarily reflect those of The Transplantation Society and/or its Sections.
  2. The hosting of material on The Transplantation Society site does not signify endorsement of this material by The Transplantation Society and/or its Sections.
  3. The material is solely for educational purposes for qualified health care professionals.
  4. The Transplantation Society and/or its Sections are not liable for any decision made or action taken based on the information contained in the material on this site.
  5. The information cannot be used as a substitute for professional care.
  6. The information does not represent a standard of care.
  7. No physician-patient relationship is being established.

Social

Contact

Staff Directory
+1-514-874-1717
info@tts.org

Address

The Transplantation Society
International Headquarters
740 Notre-Dame Ouest
Suite 1245
Montréal, QC, H3C 3X6
Canada