Differentiation of Human Endometrial Stem Cells into Schwann Cells in Fibrin Hydrogel as 3D Culture

Abstract

Human endometrial stem cells (hEnSCs) are a new source of adult multipotent stem cells with the ability of differentiation into many cell lineages. Many stem cell sources are desirable for differentiation into Schwann cells. Schwann-like cells derived from hEnSCs may be one of the ideal alternative cell sources for Schwann cell generation. In this study, for differentiation of hEnSCs into Schwann cells, hEnSCs were induced with RA/FSK/PDGF-AA/HRG as an induction medium for 14 days. The cells were cultured in a tissue culture plate (TCP) and fibrin gel matrix. The viability of cultured cells in the fibrin gel and TCP was analyzed with 3-[4,5-dimethyl-2-thia-zolyl]-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay for 7 days. The attachment of cells was analyzed with SEM and DAPI staining. The expression of S100 and P75 as Schwann cell markers was evaluated by immunocytochemistry and quantitative real-time PCR (RT-PCR). The evaluation of the MTT assay and gene expression showed that the survival rate and differentiation of hEnSCs into Schwann cells in the fibrin gel were better than those in the TCP group. These results suggest that human EnSCs can be differentiated into Schwann cells in the fibrin gel better than in the TCP, and the fibrin gel might provide a suitable three-dimensional (3D) scaffold for clinical applications for cell therapy of the nervous system.

Endometrial Stem Cell Markers: Current Concepts and Unresolved Questions

“…Endometrial stromal ASCs were also shown to be capable of multi-lineage differentiation into fat, bone (confirmed with presence of osteopontin, ostenectin and alkaline phosphatase) [ 22 ], cartilage, skeletal muscle [ 23 , 24 ], and smooth muscle (expressing specific smooth muscle cell markers including alpha-smooth muscle actin (α-SMA), desmin, vinculin and calponin) [ 25 ]. Plasticity has also been shown by the trans-differentiation of endometrial stromal ASCs into neural (neural and glial lineage markers such as Nestin, NF-L, MAP2, PDGFRa, CNP, Olig2, MBP and GFAP) [ 24 , 26 ], Schwann cells (expression of S100 and P75 noted) [ 27 ], Oligodendrocytes [ 28 ], pancreatic cells (shown by secretion of insulin and markers of β cells such as PDX1, proinsulin and c-peptide) [ 29 , 30 ], urinary bladder epithelial cells (urothelium, as tested by urothelium-specific genes and proteins, uroplakin-Ia/Ib, II, III and cytokeratin 20) [ 31 ], hepatocytes (biomarkers albumin and cytokeratin 8, reduced α-fetoprotein and α-SMA expression, synthesised urea, and stored glycogen) [ 32 , 33 ], and megakaryocytes (identified by expression of CD41a and CD42b and reduction of pluripotent transcription factors Oct4 and Sox2, platelets were seen as functional as evidenced by the upregulation of CD62p expression and fibrinogen binding following thrombin stimulation), both in vitro and in animal models [ 34 , 35 ].…”

Fibrin gel as a scaffold for photoreceptor cells differentiation from conjunctiva mesenchymal stem cells in retina tissue engineering

“…Stem cells have important role in regenerative medicine but the viability of these cells after transplantation into injured tissue are very low, so for solving this problem, we can use materials as a vehicle for enhancement of cell viability, promotion of the nutrient diffusion, soluble factors and metabolites. Understanding of the scaffold properties is very important for production of a new functional matrix in cells can regenerate the desired tissue structures [11,36]. In the present study, we examined the influence of fibrin gel on differentiation of CJMSCs to photoreceptor like cell lineages.…”

“…It is used as a biodegradable and biocompatible injection scaffold for tissue engineering. Furthermore, it has no toxic effects and its physical properties are easily adjusted to prepare a suitable environment for photoreceptor cells [11,23,25,36]. Fibrin gel was used as a scaffold, which is biodegradable, has no cytotoxicity, and is biocompatible.…”

The endometrium as a source of mesenchymal stem cells in domestic animals and possible applications in veterinary medicine

“…Apart from the common mesodermal lineages, endometrial MSCs may be induced to differentiate in vitro into skeletal and cardiac muscle, 86,87 insulin-producing cells 88 and the urinary epithelium 89 in the endodermic lineages and finally into neuronal cells [similar to neurolemmocytes, Schwann cells], 90,91 and dopaminergic neurons 91 in the ectodermic lineages. 28,92,93 Therefore, these cells might be used to treat affections such as cardiac disease, muscle degeneration, diabetes type 1 and neurodegenerative diseases in the future.…”