scholarly journals Natural Membrane Differentiates Human Adipose-Derived Mesenchymal Stem Cells to Neurospheres by Mechanotransduction Related to YAP and AMOT Proteins

Membranes ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 687
Author(s):  
Nathalia Barth de Oliveira ◽  
Ana Carolina Irioda ◽  
Priscila Elias Ferreira Stricker ◽  
Bassam Felipe Mogharbel ◽  
Nádia Nascimento da Rosa ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Neuronal Differentiation ◽  
Rt Pcr ◽  
Isolated Cells ◽  
Neuronal Markers ◽  
Enzymatic Dissociation ◽  
Trilineage Differentiation ◽  
Good Cell

Adipose tissue-derived mesenchymal stem cells (ADMSCs) are promising candidates for regenerative medicine, as they have good cell yield and can differentiate into several cell lines. When induced to the neuronal differentiation, they form neurospheres composed of neural precursors (NPs) that can be an alternative in treating neurodegenerative diseases. This study aimed to characterize NPs from neurospheres obtained after seeding ADMSCs on a natural polyisoprene-based membrane. The ADMSCs were isolated from adipose tissue by enzymatic dissociation, were subjected to trilineage differentiation, and were characterized by flow cytometry for specific ADMSC surface markers. For neuronal differentiation, the cells were seeded on polystyrene flasks coated with the membrane and were characterized by immunocytochemistry and RT-PCR. The results demonstrated that the isolated cells showed characteristics of ADMSCs. At 15 to 25 days, ADMSCs seeded on the natural membrane developed neurospheres. Then, after dissociation, the cells demonstrated characteristic neuronal markers expressed on NPs: nestin, ß-III tubulin, GFAP, NeuN, and the YAP1/AMOT in the cytoplasm. In conclusion, it was demonstrated that this membrane differentiates the ADMSCs to NPs without any induction factors, and suggests that their differentiation mechanisms are related to mechanotransduction regulated by the YAP and AMOT proteins.

scholarly journals Characterization of neuronal precursors obtained from human adipose-derived mesenchymal stem cells

2021 ◽  
Vol 31 (Supplement_2) ◽  
Author(s):  
N B Oliveira ◽  
A C Irioda ◽  
P E F Stricker ◽  
B F Mogharbel ◽  
N N Rosa ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Neural Cells ◽  
High Plasticity ◽  
Rt Pcr ◽  
Isolated Cells ◽  
Neural Precursors ◽  
Hla Dr ◽  
Neuronal Precursors ◽  
Neural Growth

Abstract Background Mesenchymal stem cells (MSCs) can be isolated from any tissue derived from the mesoderm and have as main characteristics: high plasticity, the ability to originate mesodermal and non-mesodermal tissues, acting in the modulation of the inflammatory response, and the tissue repair. When grown in microenvironments with elasticity comparable to the human brain, these cells can differentiate efficiently in neural cells due to the mechanism related to the YAP protein, which can mediate responses to substrate stiffness in mesenchymal stem cells. Methods Human adipose-derived MSCs were isolated*, then it was done the trilineage test into adipocytes, osteocytes and, chondrocytes. Besides that, differentiation to neural precursor cells was through neurospheres after seeding the cells over a natural biopolymer matrix as NFBX. Those cells were analyzed using flow cytometry for the surface markers CD13, CD34, CD45, CD73, CD90, CD105, HLA-DR, HLA-ABC, immunocytochemistry for the proteins Nestina, ß-tubulin III, YAP and AMOT and RT-PCR for the NEFM and TUBB3 genes. Results Isolated cells demonstrated characteristics of MSCs. Those cells were differentiated in neural precursors, expressing the proteins Nestina and ß-tubulin III on immunocytochemistry and, the NEFM and TUBB3 genes in RT-PCR. Regarding the YAP and AMOT proteins, it was possible to observe the translocation of the YAP protein in response to the regulation of AMOT out of the cell nucleus, proving neurodifferentiation. Conclusions Human adipose-derived MSCs seeded in a natural biopolymer matrix were able to differentiate into neural precursors expressing characteristic neural markers without adding any neural growth factors or genetic induction.

scholarly journals Treatment of Periodontal Ligament Stem Cells with MOR and CBD Promotes Cell Survival and Neuronal Differentiation via the PI3K/Akt/mTOR Pathway

2018 ◽  
Vol 19 (8) ◽  
pp. 2341 ◽  
Author(s):  
Veronica Lanza Cariccio ◽  
Domenico Scionti ◽  
Antonio Raffa ◽  
Renato Iori ◽  
Federica Pollastro ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Neuronal Differentiation ◽  
Periodontal Ligament ◽  
Combined Treatment ◽  
Mtor Pathway ◽  
Periodontal Ligament Stem Cells ◽  
Differentiation Potential ◽  
Neuronal Markers

Periodontal ligament mesenchymal stem cells (hPDLSCs), as well as all mesenchymal stem cells, show self-renewal, clonogenicity, and multi-tissue differentiation proprieties and can represent a valid support for regenerative medicine. We treated hPDLSCs with a combination of Moringin (MOR) and Cannabidiol (CBD), in order to understand if treatment could improve their survival and their in vitro differentiation capacity. Stem cells survival is fundamental to achieve a successful therapy outcome in the re-implanted tissue of patients. Through NGS transcriptome analysis, we found that combined treatment increased hPDLSCs survival, by inhibition of apoptosis as demonstrated by enhanced expression of anti-apoptotic genes and reduction of pro-apoptotic ones. Moreover, we investigated the possible involvement of PI3K/Akt/mTOR pathway, emphasizing a differential gene expression between treated and untreated cells. Furthermore, hPDLSCs were cultured for 48 h in the presence or absence of CBD and MOR and, after confirming the cellular viability through MTT (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazoliumbromide) assay, we examined the presence of neuronal markers, through immunofluorescence analysis. We found an increased expression of Nestin and GAP43 (growth associated protein 43) in treated cells. In conclusion, hPDLSCs treated with Moringin and Cannabidiol showed an improved survival capacity and neuronal differentiation potential.

Cholinergic and dopaminergic neuronal differentiation of human adipose tissue derived mesenchymal stem cells

2017 ◽  
Vol 233 (2) ◽  
pp. 936-945 ◽  
Author(s):  
Hany El Sayed Marei ◽  
Aya El-Gamal ◽  
Asma Althani ◽  
Nahla Afifi ◽  
Ahmed Abd-Elmaksoud ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Neuronal Differentiation ◽  
Human Adipose Tissue

scholarly journals Comparison of the neuronal differentiation abilities of bone marrow-derived and adipose tissue-derived mesenchymal stem cells

2017 ◽  
Vol 16 (4) ◽  
pp. 3877-3886 ◽  
Author(s):  
Yani Zheng ◽  
Chao Huang ◽  
Fang Liu ◽  
Haiyan Lin ◽  
Xiangqun Yang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Neuronal Differentiation

scholarly journals The Different Molecular Code in Generation of Dopaminergic Neurons from Astrocytes and Mesenchymal Stem Cells

2021 ◽  
Vol 22 (22) ◽  
pp. 12141
Author(s):  
Nana Wang ◽  
Xingrui Ji ◽  
Yue Wu ◽  
Shaocong Zhou ◽  
Huiyu Peng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Transcription Factor ◽  
Adipose Tissue ◽  
Dopaminergic Neurons ◽  
Cell Replacement Therapy ◽  
Cell Replacement ◽  
Neuronal Markers ◽  
Molecular Code

Transplantation of exogenous dopaminergic (DA) neurons is an alternative strategy to replenish DA neurons that have lost along the course of Parkinson’s disease (PD). From the perspective of ethical acceptation, the source limitations, and the intrinsic features of PD pathology, astrocytes (AS) and mesenchymal stem cells (MSCs) are the two promising candidates of DA induction. In the present study, we induced AS or MSCs primary culture by the combination of the classical transcription-factor cocktails Mash1, Lmx1a, and Nurr1 (MLN), the chemical cocktails (S/C/D), and the morphogens SHH, FGF8, and FGF2 (S/F8/F2); the efficiency of induction into DA neurons was further analyzed by using immunostaining against the DA neuronal markers. AS could be efficiently converted into the DA neurons in vitro by the transcriptional regulation of MLN, and the combination with S/C/D or S/F8/F2 further increased the conversion efficiency. In contrast, MSCs from umbilical cord (UC-MSCs) or adipose tissue (AD-MSCs) showed moderate TH immunoreactivity after the induction with S/F8/F2 instead of with MLN or S/C/D. Our data demonstrated that AS and MSCs held lineage-specific molecular codes on the induction into DA neurons and highlighted the unique superiority of AS in the potential of cell replacement therapy for PD.

scholarly journals Adipose-Derived Mesenchymal Stem Cells From a Hypoxic Culture Improve Neuronal Differentiation and Nerve Repair

2021 ◽  
Vol 9 ◽  
Author(s):  
Szu-Hsien Wu ◽  
Yu-Ting Liao ◽  
Kuang-Kai Hsueh ◽  
Hui-Kuang Huang ◽  
Tung-Ming Chen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Animal Model ◽  
Mesenchymal Stem Cells ◽  
Sciatic Nerve ◽  
Neuronal Differentiation ◽  
Gastrocnemius Muscle ◽  
Differentiation Potential ◽  
Neuronal Markers

Hypoxic expansion has been demonstrated to enhance in vitro neuronal differentiation of bone-marrow derived mesenchymal stem cells (BMSCs). Whether adipose-derived mesenchymal stem cells (ADSCs) increase their neuronal differentiation potential following hypoxic expansion has been examined in the study. Real-time quantitative reverse transcription-polymerase chain reaction and immunofluorescence staining were employed to detect the expression of neuronal markers and compare the differentiation efficiency of hypoxic and normoxic ADSCs. A sciatic nerve injury animal model was used to analyze the gastrocnemius muscle weights as the outcomes of hypoxic and normoxic ADSC treatments, and sections of the regenerated nerve fibers taken from the conduits were analyzed by histological staining and immunohistochemical staining. Comparisons of the treatment effects of ADSCs and BMSCs following hypoxic expansion were also conducted in vitro and in vivo. Hypoxic expansion prior to the differentiation procedure promoted the expression of the neuronal markers in ADSC differentiated neuron-like cells. Moreover, the conduit connecting the sciatic nerve gap injected with hypoxic ADSCs showed the highest recovery rate of the gastrocnemius muscle weights in the animal model, suggesting a conceivable treatment for hypoxic ADSCs. The percentages of the regenerated myelinated fibers from the hypoxic ADSCs detected by toluidine blue staining and myelin basic protein (MBP) immunostaining were higher than those of the normoxic ones. On the other hand, hypoxic expansion increased the neuronal differentiation potential of ADSCs compared with that of the hypoxic BMSCs in vitro. The outcomes of animals treated with hypoxic ADSCs and hypoxic BMSCs showed similar results, confirming that hypoxic expansion enhances the neuronal differentiation potential of ADSCs in vitro and improves in vivo therapeutic potential.

scholarly journals Ghrelin peptide improves glial conditioned medium effects on neuronal differentiation of human adipose mesenchymal stem cells

2021 ◽  
Author(s):  
Cristina Russo ◽  
Giuliana Mannino ◽  
Martina Patanè ◽  
Nunziatina Laura Parrinello ◽  
Rosalia Pellitteri ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Conditioned Medium ◽  
Neural Differentiation ◽  
Conditioned Media ◽  
Glial Fibrillary Acid Protein ◽  
Ghrelin Receptor ◽  
Neuronal Markers ◽  
Adipose Mesenchymal Stem Cells ◽  
Marker Expression

AbstractThe influences of ghrelin on neural differentiation of adipose-derived mesenchymal stem cells (ASCs) were investigated in this study. The expression of typical neuronal markers, such as protein gene product 9.5 (PGP9.5) and Microtubule Associated Protein 2 (MAP2), as well as glial Fibrillary Acid Protein (GFAP) as a glial marker was evaluated in ASCs in different conditions. In particular, 2 µM ghrelin was added to control ASCs and to ASCs undergoing neural differentiation. For this purpose, ASCs were cultured in Conditioned Media obtained from Olfactory Ensheathing cells (OEC-CM) or from Schwann cells (SC-CM). Data on marker expression were gathered after 1 and 7 days of culture by fluorescence immunocytochemistry and flow cytometry. Results show that only weak effects were induced by the addition of only ghrelin. Instead, dynamic ghrelin-induced modifications were detected on the increased marker expression elicited by glial conditioned media. In fact, the combination of ghrelin and conditioned media consistently induced a further increase of PGP9.5 and MAP2 expression, especially after 7 days of treatment. The combination of ghrelin with SC-CM produced the most evident effects. Weak or no modifications were found on conditioned medium-induced GFAP increases. Observations on the ghrelin receptor indicate that its expression in control ASCs, virtually unchanged by the addition of only ghrelin, was considerably increased by CM treatment. These increases were enhanced by combining ghrelin and CM treatment, especially at 7 days. Overall, it can be assumed that ghrelin favors a neuronal rather than a glial ASC differentiation.

scholarly journals Comparative analysis of mouse bone marrow and adipose tissue mesenchymal stem cells for critical limb ischemia cell therapy

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Pegah Nammian ◽  
Seyedeh-Leili Asadi-Yousefabad ◽  
Sajad Daneshi ◽  
Mohammad Hasan Sheikhha ◽  
Seyed Mohammad Bagher Tabei ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Cell Therapy ◽  
Femoral Artery ◽  
Critical Limb Ischemia ◽  
Limb Ischemia

Abstract Introduction Critical limb ischemia (CLI) is the most advanced form of peripheral arterial disease (PAD) characterized by ischemic rest pain and non-healing ulcers. Currently, the standard therapy for CLI is the surgical reconstruction and endovascular therapy or limb amputation for patients with no treatment options. Neovasculogenesis induced by mesenchymal stem cells (MSCs) therapy is a promising approach to improve CLI. Owing to their angiogenic and immunomodulatory potential, MSCs are perfect candidates for the treatment of CLI. The purpose of this study was to determine and compare the in vitro and in vivo effects of allogeneic bone marrow mesenchymal stem cells (BM-MSCs) and adipose tissue mesenchymal stem cells (AT-MSCs) on CLI treatment. Methods For the first step, BM-MSCs and AT-MSCs were isolated and characterized for the characteristic MSC phenotypes. Then, femoral artery ligation and total excision of the femoral artery were performed on C57BL/6 mice to create a CLI model. The cells were evaluated for their in vitro and in vivo biological characteristics for CLI cell therapy. In order to determine these characteristics, the following tests were performed: morphology, flow cytometry, differentiation to osteocyte and adipocyte, wound healing assay, and behavioral tests including Tarlov, Ischemia, Modified ischemia, Function and the grade of limb necrosis scores, donor cell survival assay, and histological analysis. Results Our cellular and functional tests indicated that during 28 days after cell transplantation, BM-MSCs had a great effect on endothelial cell migration, muscle restructure, functional improvements, and neovascularization in ischemic tissues compared with AT-MSCs and control groups. Conclusions Allogeneic BM-MSC transplantation resulted in a more effective recovery from critical limb ischemia compared to AT-MSCs transplantation. In fact, BM-MSC transplantation could be considered as a promising therapy for diseases with insufficient angiogenesis including hindlimb ischemia.

Sign in / Sign up

Export Citation Format

Share Document