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.

Isolation and Characterization of Mesenchymal Stem Cells from Canine Ovarian Surface Epithelium

2021 ◽  
Author(s):  
Ajeet Kumar Jha ◽  
Anirban Mandal ◽  
Kalyani Ray ◽  
Shyamal Kanti Guha
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stromal Vascular Fraction ◽  
Ovarian Surface Epithelium ◽  
Surface Epithelium ◽  
High Plasticity ◽  
Rt Pcr ◽  
Differentiation Potential ◽  
Ovarian Surface ◽  
Isolation And Characterization

Background: Few studies have confirmed the presence of ovarian tissue stem cells indicating the capacity for differentiation. Based on this fact, it was hypothesized that mesenchymal stem cells (MSC) were found in ovarian surface epithelium (OSE) of canines that could easily be isolated. Methods: Both left and right ovaries were minced and digested using collagenase to obtain a stromal vascular fraction (SVF). MSCs were characterized using RT-PCR. To ascertain the trilineage differentiation potential, MSCs were stained with respective stain for osteocytes, chondrocytes and adipocytes. Result: We observed elongated, spindle-shaped and fibroblast like appearance of cells after 72 h of initial culture. Expression of MSC specific surface markers were observed through RT-PCR. Using Stem Pro® differentiation medium, OSE were differentiated into osteogenic, chondrogenic and adipogenic lineages and were found to be potential source for isolation, characterization and differentiation of MSCs. Canine (OSE) is easily accessible, multipotent and has high plasticity, holding promise for applications in regenerative medicine.

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 ISOLATION OF AMNIOTIC FLUID MESENCHYMAL STEM CELLS (AF-MSCs) OBTAINED FROM CAESAREAN SECTIONS

2019 ◽  
Vol 2 (1) ◽  
pp. 1-9
Author(s):  
Bobby Indra Utama
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Amniotic Fluid ◽  
Pelvic Organ ◽  
Rt Pcr ◽  
Third Trimester ◽  
Amniotic Cavity ◽  
Hla Dr ◽  
Embryonic Antigen ◽  
Stage Specific Embryonic Antigen

Amniotic fluid is a liquid that fills the amniotic cavity which has defense and nutritional functions in fetal development. Human aterm amniotic fluid can be an ideal alternative as a source of mesenchymal stem cells, originating from the neonate. Preclinical studies of second and third trimester amnion fluid cells confirmed the number of potential donors from this wasted material. In several studies, AF-MSCs express mesenchymal markers such as CD90, CD73 (SH3, SH), CD105 (SH2), CD29, CD166, CD49e, CD58 and CD44 (MHC class I). These cells also express HLA-ABC antigens, CD 34, CD 45 which are hematopoietic markers, and endothelial CD31 markers. There is no expression of CD10, CD11b, CD14, CD34, CD117, EMA and HLA-DR, DP, DQ antigens. Most of AF-MSCs have pluripotent properties which are characterized by the discovery of octamer binding protein 3/4 (Oct-3/4), transcription factors Nanog (Nanog), and stage-specific embryonic antigen 4 (SSEA-4) on RT-PCR examination. From this study, 8 million cells was isolated. These cells will be used for research on pelvic organ prolapse therapy by using AF-MSCs. AF-MSCs isolation totally takes 6 weeks. From 1 flask, 2 million of stem cells was obtained. Keywords: amniotic fluid, AF-MSCs

scholarly journals ISOLATION OF AMNIOTIC FLUID MESENCHYMAL STEM CELLS (AF-MSCs) OBTAINED FROM CAESAREAN SECTIONS

2019 ◽  
Vol 2 (1) ◽  
pp. 1-9
Author(s):  
Bobby Indra Utama
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Amniotic Fluid ◽  
Pelvic Organ ◽  
Rt Pcr ◽  
Third Trimester ◽  
Amniotic Cavity ◽  
Hla Dr ◽  
Embryonic Antigen ◽  
Stage Specific Embryonic Antigen

Amniotic fluid is a liquid that fills the amniotic cavity which has defense and nutritional functions in fetal development. Human aterm amniotic fluid can be an ideal alternative as a source of mesenchymal stem cells, originating from the neonate. Preclinical studies of second and third trimester amnion fluid cells confirmed the number of potential donors from this wasted material. In several studies, AF-MSCs express mesenchymal markers such as CD90, CD73 (SH3, SH), CD105 (SH2), CD29, CD166, CD49e, CD58 and CD44 (MHC class I). These cells also express HLA-ABC antigens, CD 34, CD 45 which are hematopoietic markers, and endothelial CD31 markers. There is no expression of CD10, CD11b, CD14, CD34, CD117, EMA and HLA-DR, DP, DQ antigens. Most of AF-MSCs have pluripotent properties which are characterized by the discovery of octamer binding protein 3/4 (Oct-3/4), transcription factors Nanog (Nanog), and stage-specific embryonic antigen 4 (SSEA-4) on RT-PCR examination. From this study, 8 million cells was isolated. These cells will be used for research on pelvic organ prolapse therapy by using AF-MSCs. AF-MSCs isolation totally takes 6 weeks. From 1 flask, 2 million of stem cells was obtained.

scholarly journals Dental Pulp-Derived Mesenchymal Stem Cells for Modeling Genetic Disorders

2021 ◽  
Vol 22 (5) ◽  
pp. 2269
Author(s):  
Keiji Masuda ◽  
Xu Han ◽  
Hiroki Kato ◽  
Hiroshi Sato ◽  
Yu Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Dental Pulp ◽  
Genetic Disorders ◽  
In Vitro Models ◽  
High Plasticity ◽  
Human Teeth ◽  
Clinical Diagnoses ◽  
Models Of Disease

A subpopulation of mesenchymal stem cells, developmentally derived from multipotent neural crest cells that form multiple facial tissues, resides within the dental pulp of human teeth. These stem cells show high proliferative capacity in vitro and are multipotent, including adipogenic, myogenic, osteogenic, chondrogenic, and neurogenic potential. Teeth containing viable cells are harvested via minimally invasive procedures, based on various clinical diagnoses, but then usually discarded as medical waste, indicating the relatively low ethical considerations to reuse these cells for medical applications. Previous studies have demonstrated that stem cells derived from healthy subjects are an excellent source for cell-based medicine, tissue regeneration, and bioengineering. Furthermore, stem cells donated by patients affected by genetic disorders can serve as in vitro models of disease-specific genetic variants, indicating additional applications of these stem cells with high plasticity. This review discusses the benefits, limitations, and perspectives of patient-derived dental pulp stem cells as alternatives that may complement other excellent, yet incomplete stem cell models, such as induced pluripotent stem cells, together with our recent data.

Neural Precursors Derived from Embryonic and Mesenchymal Stem Cells Ameliorate Parkinson's Disease Associated Motor Impairment

2019 ◽  
Vol 254 ◽  
pp. 148
Author(s):  
Nazaret Gamez Ruiz ◽  
George A Edwards ◽  
Priyadarshini Peter ◽  
Enrique Antonio Armijo Fuentes ◽  
Carlos Kramm Barria ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Stem Cells ◽  
Parkinson's Disease ◽  
Mesenchymal Stem Cells ◽  
Motor Impairment ◽  
Neural Precursors

Mesenchymal stem cells in infantile haemangioma

2011 ◽  
Vol 64 (3) ◽  
pp. 232-236 ◽  
Author(s):  
Tinte Itinteang ◽  
Anasuya Vishvanath ◽  
Darren J Day ◽  
Swee T Tan
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Neural Crest ◽  
Capillary Endothelium ◽  
Fatty Tissue ◽  
Rt Pcr ◽  
Functional Potential ◽  
Infantile Haemangioma ◽  
Mesenchymal Differentiation ◽  
Associated Proteins

BackgroundFibro-fatty deposition commonly occurs during involution of infantile haemangioma (IH). Mesenchymal stem cells have been identified in this tumour and have been proposed to be recruited from the bone marrow and/or adjacent niches, and then give rise to the fibro-fatty tissue. The authors have recently demonstrated that the capillary endothelium of proliferating IH co-expresses primitive mesodermal, mesenchymal and neural crest markers and proposed that this same endothelium has the ability to give rise to cells of mesenchymal lineage that constitute the fibro-fatty deposition.MethodsImmunohistochemistry and real-time RT-PCR were used to further characterise proliferating IHs and haemangioma explant-derived cells (HaemEDCs).ResultsThe authors have further confirmed expression of the mesenchymal-associated proteins including preadipocyte factor-1, a mesenchymal differentiation inhibition-associated cytokine. The HaemEDCs could be differentiated into osteoblasts and adipocytes, indicating their functional potential for terminal differentiation.DiscussionThe collective expression of neural crest, mesenchymal and mesenchymal differentiation inhibition-associated proteins on the endothelium of proliferating IH suggests that the cells in the capillary endothelium within the lesion possess the ability to undergo terminal mesenchymal differentiation during the proliferating phase, but are inhibited from doing so.

218 DONOR-MATCHED FUNCTIONAL AND MOLECULAR CHARACTERIZATION OF CANINE MESENCHYMAL STEM CELLS DERIVED FROM DIFFERENT ORIGINS

2012 ◽  
Vol 24 (1) ◽  
pp. 221
Author(s):  
S. A. Ock ◽  
G. H. Maeng ◽  
Y. M. Lee ◽  
T. H. Kim ◽  
B. M. Kumar ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Transcription Factors ◽  
Telomere Length ◽  
Telomerase Activity ◽  
Rt Pcr ◽  
Cytochemical Staining ◽  
Single Donor ◽  
Cell Capacity

Canine mesenchymal stem cells (cMSC) have been successfully isolated from several adult tissue sources. However, differences in the biological properties of MSC have been shown to be associated with donor variability. Further, the stem cell capacity of cMSC of various tissues isolated from a single donor is currently unclear. Therefore, this study investigated the functional and molecular characteristics of cMSC derived from bone marrow (cBM-MSC), adipose tissue (cA-MSC) and dermal skin (cDS-MSC) of a single donor. Three kinds of cMSC were isolated by following previously published protocols. AP activity was assessed with a chromogen kit (Abcam Inc., Cambridge, MA, USA). Expression of CD markers (CD45, 90 and 105) and stem cell transcription factors (Oct3/4, Nanog and Sox2) was analysed by immunocytochemical staining. All cells were induced into osteogenesis and adipogenesis by following protocols described earlier and confirmed by cytochemical staining and the detection of lineage specific genes by RT-PCR. Chromosomal stability was assessed by a method described earlier (Ock and Rho 2008 J. Vet. Med. Sci. 70, 1165–1172) and cell cycle status was determined by a flow cytometry. Telomere length was analysed by Telo TAGGG Telomere Length Assay kit (Roche, Mannheim, Germany) and telomerase activity was evaluated by semiquantitative nested RT-PCR. Statistical analysis was performed by ANOVA using SPSS 12.0 and significance was tested when P < 0.05. Expressions of AP activity and the transcription factors, such as Oct3/4, Nanog and Sox2 were absent in all cMSC. All 3 types of cMSC positively expressed the surface markers CD90 and 105 but not CD45. Exposure of all cell lines to osteogenic and adipogenic induction medium resulted in the calcium deposition evidenced by Alizarin red S staining and the accumulation of fat globules indicated by Oil red O staining, respectively. Differentiation was further confirmed by the detection of marker genes, such as Runx2 and Pparγ. However, the degree of osteogenic or adipogenic differentiation among the 3 kinds of cMSC was different and particularly, cA-MSC had enhanced cytochemical staining associated with expression of specific genes, Runx2 and Pparγ. Ploidy analysis showed that the diploid rate was high with over 90% in all cMSC and indicated no noticeable chromosomal abnormalities. Further, less than 52% of cells were found at G1 phase in all cMSC, with lowest percentage observed in cDS-MSC (33.3%). Regardless of varied tissue sources, cMSC from a single donor showed no differences in telomere lengths (∼18–19 kbp), but the telomerase activity was different with significantly higher levels found in cBM-MSC. In conclusion, the above results suggest that tissue specific cMSC derived from a single donor possess differences in stem cell capacity and support the consideration of tissue source before judging the suitability of cells for therapeutic applications. This work was supported by grant from Basic Science Research Program through NRF funded by the Ministry of Education, Science and Technology (2009-0064229).

296 IN VITRO DIFFERENTIATION OF PORCINE BONE MARROW-DERIVED MESENCHYMAL STEM CELLS INTO HEPATOCYTE-LIKE CELLS

2013 ◽  
Vol 25 (1) ◽  
pp. 295
Author(s):  
B. Mohana Kumar ◽  
W. J. Lee ◽  
Y. M. Lee ◽  
R. Patil ◽  
S. L. Lee ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Class Ii ◽  
In Vitro Differentiation ◽  
Rt Pcr ◽  
Hepatic Differentiation ◽  
Alizarin Red ◽  
Differentiation Potential

Mesenchymal stem cells (MSC) are isolated from bone marrow or other tissues, and have properties of self renewal and multilineage differentiation ability. The current study investigated the in vitro differentiation potential of porcine bone marrow derived MSCs into hepatocyte-like cells. The MSC were isolated from the bone marrow of adult miniature pigs (7 months old, T-type, PWG Micro-pig®, PWG Genetics, Seoul, Korea) and adherent cells with fibroblast-like morphology were cultured on plastic. Isolated MSCs were positive for CD29, CD44, CD73, CD90, and vimentin, and negative for CD34, CD45, major histocompatibility complex-class II (MHC-class II), and swine leukocyte antigen-DR (SLA-DR) by flow cytometry analysis. Further, trilineage differentiation of MSC into osteocytes (alkaline phosphatase, von Kossa and Alizarin red), adipocytes (Oil Red O), and chondrocytes (Alcian blue) was confirmed. Differentiation of MSC into hepatocyte-like cells was induced with sequential supplementation of growth factors, cytokines, and hormones for 21 days as described previously (Taléns-Visconti et al. 2006 World J. Gastroenterol. 12, 5834–5845). Morphological analysis, expression of liver-specific markers, and functional assays were performed to evaluate the hepatic differentiation of MSC. Under hepatogenic conditions, MSC acquired cuboidal morphology with cytoplasmic granules. These hepatocyte-like cells expressed α-fetoprotein (AFP), albumin (ALB), cytokeratin 18 (CK18), cytochrome P450 7A1 (CYP7A1), and hepatocyte nuclear factor 1 (HNF-1) markers by immunofluorescence assay. In addition, the expression of selected markers was demonstrated by Western blotting analysis. In accordance with these features, RT-PCR revealed transcripts of AFP, ALB, CK18, CYP7A1, and HNF-1α. Further, the relative expression levels of these transcripts were analysed by quantitative RT-PCR after normalizing to the expression of the endogenous control, glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Data were analysed statistically by one-way ANOVA using PASW statistics 18 (SPSS Inc., Chicago, IL, USA), and significance was considered at P < 0.05. The results showed that the relative expressions of selected marker genes in hepatocyte-like cells were significantly increased compared with that in untreated MSC. The generated hepatocyte-like cells showed glycogen storage as analysed by periodic acid-Schiff (PAS) staining. Moreover, the induced cells produced urea at Day 21 of culture compared with control MSC. In conclusion, our results indicate the potential of porcine MSC to differentiate in vitro into hepatocyte-like cells. Further studies on the functional properties of hepatocyte-like cells are needed to use porcine MSC as an ideal source for liver cell therapy and preclinical drug evaluation. This work was supported by Basic Science Research Program through the National Research Foundation (NRF), funded by the Ministry of Education, Science and Technology (2010-0010528) and the Next-Generation BioGreen 21 Program (No. PJ009021), Rural Development Administration, Republic of Korea.

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