Isolation of Human Amnion Epithelial Cells According to Current Good Manufacturing Procedures

Abstract

Different cell types can be isolated from human placental tissues, and some have been reported to retain phenotypic plasticity and characteristics that make them a promising source of cells for regenerative medicine. Among these are human amnion epithelial cells (hAECs). Adoption of current good manufacturing practices (cGMP) and enhanced quality control is essential when isolating hAECs in order to deliver a safe and effective cellular product for clinical purposes. This unit describes a detailed protocol for selective isolation of hAECs from human term placenta with little to no contamination by other cell types. A method for characterizing the heterogeneity of the hAEC suspension is also provided. The resulting cell product will be useful for clinical as well as basic research applications. © 2016 by John Wiley & Sons, Inc.

Ectonucleotidase Expression on Human Amnion Epithelial Cells: Adenosinergic Pathways and Dichotomic Effects on Immune Effector Cell Populations

“…Discarded placentae from uncomplicated cesarean resections at 37-42 wk of gestational age were obtained from Karolinska University Hospital (Stockholm, Sweden) after the approval by the Institutional Review Board. Placental hAEC were isolated as previously described (4). Briefly, amnion membrane was surgically removed from the surface of the placenta and washed to remove blood cells.…”

“…Briefly, amnion membrane was surgically removed from the surface of the placenta and washed to remove blood cells. TrypLE (Life Technologies) digestion (30 min) was specifically performed to release epithelial cells without altering cell surface Ags, as previously described (3,4). Viability after isolation ranged from 90 to 97%, whereas after cryopreservation it ranged from 85 to 94%.…”

“…To our knowledge, our group was the first to identify and describe the nature of stem cells of human amnion epithelial cells (hAEC) (3). We developed efficient techniques to isolate hAEC with epithelial characteristics and a heterogeneous expression of membrane surface Ags (4). The hAEC phenotype includes classic epithelial adhesion molecules, such as EpCAM (CD325) and integrin subunits (CD29 and CD49f).…”

“…The hAEC phenotype includes classic epithelial adhesion molecules, such as EpCAM (CD325) and integrin subunits (CD29 and CD49f). On the contrary, markers routinely used to define and identify stromal cells are not detectable (4,5). The surface markers commonly observed on embryonic stem cells (e.g., globoseries glycolipids SSEA-3 and -4 and keratansulfateassociated Ags TRA 1-60 and 1-81) are consistently expressed, perhaps contributing to their multilineage differentiation potential (1,3,6).…”

Producing Human Amniotic Epithelial Cells-only Membrane for Transplantation

“…Then, these hAECs could happen epithelial-to-mesenchymal transition, or spontaneous differentiation, which were adverse to their clinical application. To address these problems, some studies used different factors such as progesterone, EGF or Supplement S7in hAECs’ culture [5,15-19,21,39,48]. However, these results were still not ideal.…”