Explant culture and multilineage differentiation of amniotic membrane derived stem cells

Abstract

Amniotic membrane derived stem cells (AMSCs) are reported to have a comparatively higher potency than multipotent stem cells. These cells are shown to have low immunogenicity and no teratogenicity. Among various conventional methods of isolation using enzymes, explant culture method is believed to be an easy and costeffective way to harvest stem cells. The purpose of this study was to isolate AMSCs from amniotic membrane of rats and to characterize them for multilineage differentiation, including generation of neurospheres to use them later in in-vivo experiments. Amniotic membranes were collected from Wistar rats on 17 th day of pregnancy. After processing of the tissues, AMSCs were isolated by the explant culture method and continued to grow until 10 th passage. The doubling time was estimated and the cells were analyzed for growth curve parameters at passages 5 and 9. The osteogenic and adipogenic differentiation studies were carried out from the same cells after 3 rd passage.Neurospheres generation from AMSCs was performed using neurogenic induction media.The cells were further assessed for their mesenchymal, haemopoietic, and neurogenic marker expressions by immunofluorescence staining and PCR analysis The study suggests that AMSCs isolated through explant culture are reliable stem cells which could generate neurospheres under proper induction conditions and could be a potential candidate to be used on in-vivo neural degeneration models.

Exploring the therapeutic potential of allogeneic amniotic membrane for quality wound healing in rabbit model

“…Using the explant culture approach, rAM-MSCs were isolated. 16 The initial culture had a heterogeneous population of cells of foetal origin, including round, polygonal and fibroblastic cells, which was in accordance with a previous research study. 16 The fibroblastic cells increased with the number of passages and a homogenous population of fibroblastic morphology cells were obtained by the 3 rd to 4 th passage.…”

“…Mature AMs were collected from healthy female rabbits following the spontaneous birth of kits which is favourable, 15 as they contain high levels of hyaluronan compared to those acquired from elective caesareans. Using the explant culture approach, rAM‐MSCs were isolated 16 . The initial culture had a heterogeneous population of cells of foetal origin, including round, polygonal and fibroblastic cells, which was in accordance with a previous research study 16 .…”