In vitrodifferentiation of human amniotic epithelial cells into insulin-producing 3D spheroids

Abstract

Regenerative medicine and stem cell therapy may represent the solution for the treatment of non-curable human diseases such as type 1 diabetes. In this context of growing demand for functional and safe stem cells, human amniotic epithelial cells (hAECs) from term placenta have attracted increasing interest for their wide availability, stem cell properties, and differentiation plasticity, which make them a promising tool for stem cell-based therapeutic applications. We initially assayed the stemness characteristics of hAECs in serum-free conditions. Subsequently we developed a culture procedure on extracellular matrix for the formation of three-dimensional (3D) spheroids. Finally, we tested the immunomodulation and differentiation potential of hAEC spheroids: the presence of pancreatic endocrine hormones was revealed with transmission electron microscopy and immunofluorescence analyses; the release of C-peptide in hyperglycemic conditions was assayed with ELISA.The serum-free culture conditions we applied proved to maintain the basic stemness characteristics of hAECs. We also demonstrated that 3D spheroids formed by hAECs in extracellular matrix can be induced to differentiate into insulin-producing cells. Finally, we proved that control and induced cells equally inhibit the proliferation of activated mononuclear cells.The results of this study highlight the properties of amnion derived epithelial cells as promising and abundant source for cell-based therapies. In particular we are the first group to show the in vitro pancreatic induction of hAECs cultured on extracellular matrix in a 3D fashion. We accordingly propose the outcomes of this study as a novel contribution to the development of future cell replacement therapies involving placenta-derived cells.

Concise Review: Fetal Membranes in Regenerative Medicine: New Tricks from an Old Dog?

“…Transplantation of hAMSC differentiated into pancreatic islet cells, into streptozotocin‐induced type I diabetic mice restored bodyweights and normalized hyperglycemia for 7 months . Encapsulation of hAMSC differentiated into insulin producing islet cells in polyurethane‐polyvinyl pyrrolidone macrocapsules have also previously been shown to protect the transplanted cells against immune rejection, while normalizing hyperglycemia in diabetic mice . Transplantation of hAMSC derived islet cells into the kidneys of mice with streptozotocin‐induced diabetes restored body weight, and normalized the blood glucose levels, which lasted for 210 days.…”

“…In a rat model of PD with 6-hydroxydopamine lesions, a higher survival rate of dopaminergic neurons was reported, along with an increase in dopaminergic neurons in the substantia nigra and restrained stem cells growth [75]. Transplantation of hAEC into a rat model of spinal cord injury significantly reduced mechanical allodynia [76], suggesting that hAEC transplantation hCMSC [66] Myocardial infarction hAMSC [63][64][65] hCMSC [67] Stroke hAEC [68,69] Neurological Traumatic brain injury hAMSC [71] Spinal cord injury hAMSC [72] hAEC [76] Optic nerve crush injury hCMSC [73] Encephalomyelitis hAEC [74] Parkinson's Disease hAEC [75] Diabetes hAEC [77] hAMSC [78,79] hCMSC [80] Gastrointestinal Proctitis hAMSC [81] Colitis hAMSC [82] liver fibrosis hCMSC [83] hAEC [84,85] Respiratory Pulmonary fibrosis hAEC [49,50,108] hAMSC, hCMSC [87] Chronic obstructive pulmonary disease hAEC [91] Asthma hAEC [92] Bronchopulmonary dysplasia hAEC [93,94] hCMSC [95] Cystic fibrosis hAMSC [96] hAEC [97] Abbreviations: hAEC, human amnion epithelial cells; hAMSC, human amnion mesenchymal stem cells; hCMSCs, human chorionic mesenchymal stem cells. [76].…”

“…A culture protocol was recently developed to grow pancreatic hormone releasing 3D spheroids of hAEC that released C-peptide in hyperglycemic conditions [77]. This may serve as a bridge to clinically useful applications of fetal membrane stem and stem-like cells, which have been shown in experimental models of diabetes to show promise.…”

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.…”

“…It’s proven that hAECs have a comprehensive pluripotency [1-3]. For instance, hAECs are capable of differentiation into osteoblasts [4], insulin-producing 3D spheroids [5], granulosa cells [6], corneal epithelial cells [7], hepatic cells [8], progenitor of cortical neurons [9], and pancreatic lineage [10]. hAECs have a characteristic of immune privilege due to a low expression of surface marker major histocompatibility complex, class II, DR (HLA-DR) [11].…”