Differentiation of Amniotic Membrane Mesenchymal Stem Cells to Cardiomyocytes and its Characteristics

Introduction:  Mesenchymal stem cells (MSCs) have high potential in regenerative medicine based on their renewal properties and multilinearity differentiation capacity. MSCs have the ability to differentiate to osteoblasts, adipocytes, chondrocytes, cardiomyocytes, nerve cells, and fibroblasts. These cells have many sources such as bone marrow, umbilical cord blood, and amniotic membrane. Amniotic membrane is a postnatal organ, which does not require an invasive method for procurement. The immunomodulatory properties of MSCs make these cells the primary choice for allotransplantation and xenotransplantation.Materials and Methods:   In this study, MSCs were isolated from the amniotic membrane, and their surface markers were identified using flow cytometry. The cells were differentiated to osteoblasts, adipocytes and cardiomyocytes using differentiation medium. GAP-43 and α-actin were studied with immunofluorescence and the expressions of related genes (GATA-4 and C-TNT) were assayed by real-time polymerase chain reaction.Results: The results confirmed the differentiation of MCS to cardiomyocytes. The expression level of GATA-4 and C-TNT was higher than that of the control. The results of the present study suggested that differentiated human amniotic MSC possessed some characteristics of cardiomyocytes.Conclusion: Therefore, according to the results, the amniotic membrane is a suitable source of mesenchymal cells for differentiation into cardiomyocyte cells. 

Studying the suppressing effect of mesenchymal stem cells derived from amniotic membrane on colorectal cancer

Mesenchymal stem cell therapy is considered as a proper tool for biological activities and treatment of diseases and cancer. Here, the suppressing effect of mesenchymal stem cells derived from the amniotic membrane (AM-MSCs) on colorectal cancer (HT29 cell line) was studied. MSCs were isolated from the human amniotic membrane and identity tests were performed. AM-MSC and HT29 cells were co-cultured and MTT assay was used to determine proper inhibitory concentration. The apoptotic effect of AM-MSC on HT29 cells was studied by Acridine orange staining. Expression of CDK2 protein and the antioxidant effect of AM-MSCs on HT29 was performed by immunocyto chemistry test. The effect of MSC on the HT29 cell line migration was determined by Scratch test. The result showed that AM-MSC had proliferation-inhibitory effects and caused apoptosis of the HT29 cell line. Real Time-PCR assay showed the increase of gene expression of p53, Caspase3 in apoptosis, and p21 in tumor cell line treated with AM-MSC. AM-MSC arrested the cell cycle of tumor cells by reduction of CDK2 protein. The migration of HT29 and ROS expression levels was decreased. Altogether, AM-MSC may play a role in suppressing colorectal cancer HT29 cell line by inhibiting angiogenesis, cell cycle, and induction of apoptosis and has antioxidant properties

The effect of Mesenchymal Stem Cells of Amniotic Membrane on the Proliferation and Differentiation of Umbilical Cord Blood CD34+ cells

Introduction: Ex vivo proliferation of hematopoietic stem cells (HSCs) of umbilical cord is widely used by combination of cytokine and stromal mesenchymal stem cells (MSCs) as feeder layer due to increase the cell doses, adequately. However, numerous studies have shown that ex vivo proliferation of these cells impairs their functions, including reduced self-renewal ability, apoptosis induction, and disordered cell cycle. MSCs have different sources such as amniotic membrane with a stable karyotype and high quality because of isolation from embryonic tissues, so that they are considered as a useful source for MSCs.Materials and Methods: In this study, isolated mesenchymal cells from the amniotic membrane were used as feeders for the HSCs proliferation. Four different cultures with various conditions were used; first one containing cytokines (stem cell factor, thrombopoietin, and FMS-related tyrosine kinase 3 ligand), second one with MSCs co-cultured with the aforementioned cytokines, third medium co-cultured with MSCs without cytokines, and finally the control medium was without co-culture condition and cytokines. Expression of mRNAs of HOXB4, GATA2, BCL2, and Survivin genes was also investigated.Results: The findings showed that the expression of mRNAs of these genes decreased in culture with cytokine, solely; however the expression of these genes was significantly higher in co-cultured system with cytokine rather than just with cytokine.Conclusion: : In general, the findings of this study indicate that the derived MSCs from amniotic membrane is a good source for the proliferation of umbilical cord blood CD34+ cells”. Because these cells increase the UCB-CD34+ quantity and their preservation properties.

Differentiation of mesenchymal stem cells isolated from the amniotic membrane and umbilical cord to osteocytes and the expression of RunX2, Osteonectin, ALP genes

Mesenchymal stem cells (MSCs) are multipotent cells and able to differentiate into connective tissues such as bone, fat, cartilage, tendon, and muscle. They show to be very potent tools for tissue engineering and regenerative medicine. Several researches have shown that amniotic membrane mesenchymal stem cells (AM-MSCs) and umbilical cord mesenchymal stem cells (UCB-MSCs) are both multipotent in nature differentiating into several cell types such as adipocytes and osteoblasts. In this study, mesenchymal stem cells were derived from the human amniotic membrane (hAM-dMSCs) and umbilical cord then characterized with their surface antigens using flow cytometry. These cells differentiated to osteocyte and adipocyte in induction medium then the expression of RunX2, Osteonectin, and ALP genes were calculated by Real-Time PCR. We showed that AM-MSCs and UCB-MSCs can discriminate to osteogenic and adipogenic cells in the specific induction medium. The capability of AM-MSCs and UCB-MSCs differentiation to osteogenic cells was confirmed by enhanced expression of RUNX2, ALP and Osteonectin gene and deposition calcium shown by alzerin staning. Given the available evidence, we conclude that AM-MSCs and UCB-MSCs have suitable access, low immunization and lack of medical ethics problems are one of the appropriate sources for differentiation in to osteogenic and adipogenic cells. Also, they can be considered as good choices for treatment of mesenchymal tissue injuries and tissue engineering