Differentiation of human amniotic epithelial cells into Schwann-like cells via indirect co-culture with Schwann cells in vitro

Abstract

Human amniotic epithelial cells (hAECs) exhibit multi‑lineage differentiation ability. The present study investigated the possibility that hAECs possess the potential to differentiate into Schwann‑like cells using an invitro indirect co‑culture approach. hAECs were isolated via enzymatic digestion, and immunocytochemistry and flow cytometry were performed to identify the hAECs. The hAECs were co‑cultured with Schwann cells (SCs) to differentiate the hAECs into Schwann‑like cells via induced proximity. The expression of typical S‑100 SC markers in the co‑cultured hAECs was determined via immunocytochemistry. For the functional experiments, reverse transcription quantitative polymerase chain reaction (RT‑qPCR) was performed to measure the expression levels of nerve growth factor (NGF), brain‑derived neurotrophic factor (BDNF) and glial cell‑derived neurotrophic factor (GDNF) mRNA. In addition, neurite outgrowth was measured in PC12 cells following co‑culture with the differentiated hAECs. Subsequent to co‑culture with SCs for 21days, the hAECs exhibited spindle‑like morphology. The immunocytochemistry results revealed that the co‑cultured hAECs expressed S‑100, indicating differentiation into Schwann‑like cells. RT‑qPCR revealed that NGF, BDNF and GDNF expression was upregulated upon differentiation. The average axon length of the PC12 cells increased from 21.32±5.45 to 51.32±8.56µm subsequent to co‑culture with the differentiated hAECs. These results demonstrate that this indirect co‑culture microenvironment induced the hAECs to differentiate into Schwann‑like cells that exhibited the morphological, phenotypic and functional characteristics of SCs. Therefore, the use of differentiated hAECs that exhibit the characteristics of SCs provides a promising alternative to the present techniques used for peripheral nerve regeneration.