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 Concentrated Hypoxia-Preconditioned Adipose Mesenchymal Stem Cell-Conditioned Medium Improves Wounds Healing in Full-Thickness Skin Defect Model

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Biao Sun ◽  
Shilei Guo ◽  
Fei Xu ◽  
Bin Wang ◽  
Xiujuan Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Conditioned Medium ◽  
Skin Defect ◽  
Full Thickness ◽  
Defect Model ◽  
Full Thickness Skin ◽  
Thickness Skin ◽  
Freeze Dried ◽  
Adipose Mesenchymal Stem Cells

In recent years, the bioactive factors were utilized in exercise and athletic skin injuries. In this research, the concentrated conditioned medium of hypoxia-preconditioned adipose mesenchymal stem cells, which is rich in bioactive factor, is applied in full-thickness skin defect model to evaluate the therapeutic efficacy. Adipose mesenchymal stem cells were harvested from the abdominal subcutaneous adipose tissues. The surface markers and the potential of differentiation were analyzed. The conditioned medium of hypoxia-preconditioned stem cells was collected and freeze-dried and then applied on the rat full-thickness skin defect model, and the healing time of each group was recorded. Haematoxylin and eosin staining of skin was assessed by microscope. The characteristics of adipose mesenchymal stem cells were similar to those of other mesenchymal stem cells. The concentration of protein in freeze-dried conditioned medium in 1 mL water was about 15 times higher than in the normal condition medium. In vivo, the concentrated hypoxia-preconditioned conditioned medium can reduce the wound size and accelerate the skin wound healing. The concentrated hypoxia-preconditioned adipose mesenchymal stem cell-conditioned medium has great effect on rat model of wound healing, and it would be an ideal agent for wound care in clinical application.

scholarly journals ARPE-19 conditioned medium promotes neural differentiation of adipose-derived mesenchymal stem cells

2021 ◽  
Vol 13 (11) ◽  
pp. 1783-1796
Author(s):  
Giuliana Mannino ◽  
Martina Cristaldi ◽  
Giovanni Giurdanella ◽  
Rosario Emanuele Perrotta ◽  
Debora Lo Furno ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Conditioned Medium ◽  
Neural Differentiation

Analysis of the Effects of Low-Dose Radiation on Human Mesenchymal Stem Cells

2021 ◽  
Vol 65 (6) ◽  
pp. 5-10
Author(s):  
D. Usupzhanova ◽  
T. Astrelina ◽  
I. Kobzeva ◽  
V. Nikitina ◽  
Yu. Suchkova ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Conditioned Medium ◽  
Adaptive Response ◽  
Proliferative Activity ◽  
Human Mesenchymal Stem Cells ◽  
Conditioned Media ◽  
X Ray ◽  
Cultivation In Vitro

Purpose: The aim of the study was to study the effect of low X-ray doses on human mesenchymal stem cells (MSCs) in long-term cultivation in vitro. Material and methods: MSCs of the mucosal gum tissue of human were used. Cells were irradiated using an RUST-M1 X-ray unit (Russia) at doses of 50, 80, 100, 250 and 1000 mGy (dose rate40 mGy/min) and then cultivated according to standard methods. Immunological characteristics and viability of MSCs were evaluated on a FACSCanto II flow cytometer (Becton Dickinson CA, USA) for early and late passengers. Proliferative activity (PA) was evaluated using an xCelligence real-time cell analyzer (ACEA Biosciencs, Inc.). Results: It was shown that the proliferative activity (PA) of MSCs of the mucosal gum tissue which were irradiated at dose 50 mGy is comparable with the control group in long-term cultivation while doses of 100 and 250 mGy showed a decrease of PA. Also non-irradiated MSCs showed a significant decrease of the PA during cultivation in a conditioned medium from cells that received dose of 1000 mGy and an increase of PA during cultivation in a conditioned medium of cells that received doses of 50, 100 and 250 mGy. The cells were previously irradiated at dose 250 mGy showed adaptive response during cultivation in conditioned medium from cells that received dose of 1000 mGy. Сonclusion: The assessment of the effects of low radiation doses was focused on the bystander effect in the presented study. It was noted after adding conditioned media from irradiated cells to previously irradiated and non-irradiated MSCs. The bystander effects for low and high doses are different and their biological meaning requires further study. The phenomenon of adaptive response was shown after addition conditioned media from cells irradiated at dose 1000 mGy to pre-irradiated MSCs received a dose of 250 mGy. The obtained result leads to the conclusion that the effects of low doses can be positive. Thus, the results of study mainly correspond to the threshold nonlinear concept, according to which the effect is not proportional to the received radiation dose.

ARPE-19 conditioned medium promotes neural differentiation of adipose-derived mesenchymal stem cells

2021 ◽  
Vol 13 (11) ◽  
pp. 1784-1797
Author(s):  
Giuliana Mannino ◽  
Martina Cristaldi ◽  
Giovanni Giurdanella ◽  
Rosario Emanuele Perrotta ◽  
Debora Lo Furno ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Conditioned Medium ◽  
Neural Differentiation

Neural differentiation of human adipose-derived mesenchymal stem cells induced by glial cell conditioned media

2018 ◽  
Vol 233 (10) ◽  
pp. 7091-7100 ◽  
Author(s):  
Debora Lo Furno ◽  
Giuliana Mannino ◽  
Rosario Giuffrida ◽  
Elisa Gili ◽  
Carlo Vancheri ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Glial Cell ◽  
Neural Differentiation ◽  
Conditioned Media

Conditioned Medium of Human Adipose Mesenchymal Stem Cells Increases Wound Closure and Protects Human Astrocytes Following Scratch Assay In Vitro

2017 ◽  
Vol 55 (6) ◽  
pp. 5377-5392 ◽  
Author(s):  
Eliana Baez-Jurado ◽  
Oscar Hidalgo-Lanussa ◽  
Gina Guio-Vega ◽  
Ghulam Md Ashraf ◽  
Valentina Echeverria ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Conditioned Medium ◽  
Wound Closure ◽  
Scratch Assay ◽  
Human Astrocytes ◽  
Adipose Mesenchymal Stem Cells

scholarly journals MTA Enhances the Potential of Adipose-Derived Mesenchymal Stem Cells for Dentin–Pulp Complex Regeneration

Materials ◽  
2020 ◽  
Vol 13 (24) ◽  
pp. 5712
Author(s):  
Danial Babaki ◽  
Kagya Amoako ◽  
Ahmad Reza Bahrami ◽  
Sanam Yaghoubi ◽  
Mahdi Mirahmadi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Conditioned Medium ◽  
Mineral Trioxide Aggregate ◽  
Cell Types ◽  
Conditioned Media ◽  
Alizarin Red ◽  
Gene Markers ◽  
Alp Activity ◽  
Cell Source

The aim of the current study was to investigate the effects of mineral trioxide aggregate (MTA) on the proliferation and differentiation of human adipose-derived mesenchymal stem cells (Ad-MSCs) as a surrogate cell source in futuristic stem-cell-based endodontic therapies. Human Ad-MSCs and mesenchymal stem cells derived from bone marrow (BM-MSCs) were isolated from liposuction waste adipose tissue and femur, respectively, and the effects of MTA-conditioned media on their viability, mineralization potential, and osteo/odontogenic differentiation capacity were subsequently evaluated. Alkaline phosphatase (ALP) activity, quantitative alizarin red S staining, and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analyses were performed to investigate and compare the osteo/odontogenic induction potential of MTA on the Ad/BM-MSCs. The results of cytotoxicity assay revealed that at different concentrations, MTA-conditioned medium was not only biocompatible toward both cell types, but also capable of promoting cell proliferation. ALP activity assay showed that 0.2 mg/mL was the optimal concentration of MTA-conditioned medium for osteo/odontogenic induction in Ad/BM-MSCs. The expression of osteo/odontogenic gene markers was increased in Ad/BM-MSCs treated with 0.2 mg/mL MTA-conditioned media. Our results indicated that MTA can efficiently enhance the osteo/odontogenic potential of Ad-MSCs, and thus they can be considered as a better cell source for dentin–pulp complex regeneration. However, further investigations are required to test these potentials in animal models.

scholarly journals Neural differentiation of canine mesenchymal stem cells/multipotent mesenchymal stromal cells

2020 ◽  
Author(s):  
Sonja Prpar Mihevc ◽  
Vesna Kokondoska Grgich ◽  
Andreja Nataša Kopitar ◽  
Luka Mohorič ◽  
Gregor Majdic
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Mesenchymal Stem Cells ◽  
Glial Cells ◽  
Stromal Cells ◽  
Neural Differentiation ◽  
Neural Induction ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Induction Medium ◽  
Neuronal Markers

Abstract Background: Ability of adipose tissue-derived multipotent mesenchymal stromal cells/mesenchymal stem cells (ASCs) to differentiate in neural lineages promises progress in the field of regenerative medicine especially for replacing damaged neuronal tissue in different neurological disorders. Reprogramming of ASCs can be induced by supplying growth medium with chemical neurogenic inductors and/or specific growth factors. We investigated the neural differentiation potential of canine ASCs using several growth media (KEM, NIMa, NIMb, NIMc) containing various combinations of neurogenic inductors: B27 supplement, valproic acid, forskolin, N2-supplement and retinoic acid. First the cells were preconditioned in proliferation medium, followed by induction of neuronal differentiation. Six canine ASCs cell lines were assessed, half from female and half from male donors. The cell morphology, growth dynamics, viability were observed along with expression of neuron and astrocyte specific markers, which were assessed by immunocytochemistry and flow cytometry. Results: After 3, 6 and 9 days, elongated neural-like cells with bipolar elongations were observed and some oval cells with light nuclei appeared. After three and nine days of neural induction, differentiation into neurons and glial cells was observed. Expression of neuronal markers tubulin beta III (TUBB3), neurofilament H (NF-H) and glial fibrillary acidic protein (GFAP) was observed by immunocytochemistry. High GFAP expression (between 70 and 90% of all cells) was detected after three days of growth in neural induction medium a (NIMa) by flow cytometry, and expression of adult neuronal markers NF-H and microtubule associated protein-2 (MAP2) was detected in around 25% of cells. After nine days of ASCs differentiation a drop in expression rates of all markers was detected. There were no differences between neural differentiation of ASCs isolated from female or male dogs. Conclusions: The differentiation repertoire of canine ASCs extends beyond mesodermal lineages. Using a defined neural induction medium the canine ASCs were able to transform to neural lineages, bearing markers of neuronal and glial cells and also displayed the typical neuronal morphology. Differentiated ASCs can be a source of neural cellular lineages for regenerative therapy of nerve damage and also could be applicable for modeling of neurodegenerative diseases.

223 COMPARISON OF NEURAL DIFFERENTIATION BETWEEN RAT AND CAT MESENCHYMAL STEM CELLS UNDER CHALLENGE BY THE SAME NEUROINDUCTION AGENT

2007 ◽  
Vol 19 (1) ◽  
pp. 228
Author(s):  
G. Z. Jin ◽  
X. F. Yu ◽  
S. J. Cho ◽  
G. Y. Park ◽  
Y. J. Lee ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Neural Differentiation ◽  
Culture Media ◽  
Expression Profiles ◽  
Neural Induction ◽  
Induction Medium ◽  
Microscopy Imaging ◽  
Neuronal Markers ◽  
Post Induction

Bone marrow-derived mesenchymal stem cells (MSCs) have the potential to differentiate into a variety of cell types, including osteocytes, chondrocytes, and adipocytes. Moreover, MSCs have the capacity to differentiate into neural lineages. The present study aimed to investigate the differences between the expression profiles during neural differentiation of rat and cat MSCs. MSCs in the 4th passage, which were isolated from the femurs and tibias by standard methodology, were used in our study. Culture media was divided into pre-induction medium, which consisted of DMEM-HG, 10% fetal bovine serum, 10 ng mL-1 of basic fibroblast growth factor (bFGF), and 1 mM �-mercaptoethanol, for 24 h, and neuron induction medium, which was composed of DMEM-HG, 2% DMSO, 200 �M BHA, 25 mMKCl, 10 �M forskolin, 5 �g mL-1 insulin-transferrin-sodium selenite, and 20 ng mL-1 bFGF, for 24 h. Thereafter, cell morphology and growth traits were determined by light microscopy imaging and by examining the cell-surface antigen profile and differentiation repertoire by immunocytochemistry, respectively. Regarding the expression of 3 MSC-related surface antigens, rat cells were positive for CD18 and CD44, whereas cat cells expressed CD9 and CD44. Under proneurogenic conditions, rat neuron-like cells progressively increased at 30 min post-induction, peaked at 1 h, and gradually declined after 12 h. At 24 h after neural induction, there were still some neuron-like cells. Meanwhile, cat cells were expressed increasingly during the first hour, peaked at 2 to 3 h, were sustained for 8 h after neural induction, and then gradually declined.A few neuron-like cells remained until 24 h post-induction. In neural differentiation, rat MSCs were positive for �III-tubulin, NF-L, NF-M, NF-H, trkA, and vimentin; cat MSCs were negative for �III-tubulin and NF-H but positive for NF-L, NF-M, trkA, and vimentin. Both MSCs were negative for oligodendrocyte markers. Our results revealed that there is variation in neural differentiation sensitivity due to species type. Rat cells were more sensitive in response to neuroinduction agents, maintained their morphology for a longer time, and expressed relatively mature neuronal markers. On the contrary, the sensitivity of cat cells was weaker, and survival time was shorter compared to that of the rat cells. The cat cells expressed immature neuronal markers. The present data suggest a significant aspect of the culture of MSCs from higher-grade species. This work was supported by KOSEF (Grant ? M10525010001-05N2501-00110).

scholarly journals Neural differentiation of canine mesenchymal stem cells/multipotent mesenchymal stromal cells

2019 ◽  
Author(s):  
Sonja Prpar Mihevc ◽  
Vesna Kokondoska Grgich ◽  
Andreja Nataša Kopitar ◽  
Luka Mohorič ◽  
Gregor Majdic
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Mesenchymal Stem Cells ◽  
Glial Cells ◽  
Stromal Cells ◽  
Neural Differentiation ◽  
Neural Induction ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Induction Medium ◽  
Neuronal Markers

Abstract Background: Ability of adipose tissue-derived multipotent mesenchymal stromal cells/mesenchymal stem cells (ASCs) to differentiate in neural lineages promises progress in the field of regenerative medicine especially for replacing damaged neuronal tissue in different neurological disorders. Reprogramming of ASCs can be induced by supplying growth medium with chemical neurogenic inductors and/or specific growth factors. We investigated the neural differentiation potential of canine ASCs using several growth media (KEM, NIMa, NIMb, NIMc) containing various combinations of neurogenic inductors: B27 supplement, valproic acid, forskolin, N2-supplement and retinoic acid. First the cells were preconditioned in proliferation medium, followed by induction of neuronal differentiation. Six canine ASCs cell lines were assessed, half from female and half from male donors. The cell morphology, growth dynamics, viability were observed along with expression of neuron and astrocyte specific markers, which were assessed by immunocytochemistry and flow cytometry. Results: After 3, 6 and 9 days, elongated neural-like cells with bipolar elongations were observed and some oval cells with light nuclei appeared. After three and nine days of neural induction, differentiation into neurons and glial cells was observed. Expression of neuronal markers tubulin beta III (TUBB3), neurofilament H (NF-H) and glial fibrillary acidic protein (GFAP) was observed by immunocytochemistry. High GFAP expression (between 70 and 90% of all cells) was detected after three days of growth in neural induction medium a (NIMa) by flow cytometry, and expression of adult neuronal markers NF-H and microtubule associated protein-2 (MAP2) was detected in around 25% of cells. After nine days of ASCs differentiation a drop in expression rates of all markers was detected. There were no differences between neural differentiation of ASCs isolated from female or male dogs. Conclusions: The differentiation repertoire of canine ASCs extends beyond mesodermal lineages. Using a defined neural induction medium the canine ASCs were able to transform to neural lineages, bearing markers of neuronal and glial cells and also displayed the typical neuronal morphology. Differentiated ASCs can be a source of neural cellular lineages for regenerative therapy of nerve damage and also could be applicable for modeling of neurodegenerative diseases.

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