Int J Stem Cells.  2018 Jun;11(1):87-95. 10.15283/ijsc18001.

Large-scale Isolation, Expansion and Characterization of Human Amniotic Epithelial Cells

Affiliations
  • 1Sri Research for Tissue Engineering Pvt. Ltd, Shankara Research Centre, Rangadore Memorial Hospital, Bangalore, India. knsridhar@sr-te.com

Abstract

BACKGROUND AND OBJECTIVES
The human Amniotic epithelial cells (AME) derived from amniotic membrane of placenta have been considered as the potential fetal stem cell source with minimal or no ethical concerns and are important therapeutic tool for anti-fibrotic and regenerative therapies.
METHODS AND RESULTS
Here, we evaluated the isolation, media screening, scale-up and characterization of AME cells. The isolation, expansion of AMEs were performed by sequential passaging and growth kinetics studies. The AMEs were characterized using immunocytochemistry, immunophenotyping, In-vitro differentiation, and anti-fibrotic assays. The growth kinetics study revealed that the AME cultured in Ultraculture (UC) and DMEM knockout (DMEM-KO) have prominently higher growth rate compared to others. Overall, the AMEs cultured from 5 different media retained basic morphological characteristics and the functional characteristics.
CONCLUSIONS
Our result suggests that the AMEs can be successfully cultured in UC based complete media without losing its epithelial cell characteristics even after passaging for passage 2 (P2). However, a careful and methodical pre-clinical and clinical translation studies need to be conducted to show its safety and efficacy.

Keyword

Amniotic epithelial cells; Cell therapy; Cryopreservation; Stability; Growth kinetics; Tissue engineering

MeSH Terms

Amnion
Cell- and Tissue-Based Therapy
Cryopreservation
Epithelial Cells*
Fetal Stem Cells
Humans*
Immunohistochemistry
Immunophenotyping
Kinetics
Mass Screening
Methods
Placenta
Tissue Engineering
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