Isolation and identification of mesenchymal stem cells from human fetal pancreas

Abstract Background: Exosomes, internal proteins, lipids, and nucleic acids coated by phospholipid bilayer membranes, are one type of small extracellular vesicles, which can mediate cell-cell communication. In recent years, exosomes have gained considerable scientific interest due to their widely applied prospect in the diagnosis and therapeutics of human and animal diseases. In this study, we describe for the first time a feasible method designed to isolate and characterize exosomes from feline plasma, urine and adipose-derived mesenchymal stem cells. Results: Exosomes from feline plasma, urine and adipose-derived mesenchymal stem cells were successfully isolated by differential centrifugation. Quantification and sizing of exosomes were assessed by transmission electron microscopy, flow nano analysis and western blotting. Detected particles showed the normal size (30-100 nm) and morphology described for exosomes, as well as presence of the transmembrane protein (TSG101, CD9, CD63, and CD81) known as exosomal marker.Conclusions: The results suggest that differential centrifugation is a feasible method for isolation of exosomes from different types of feline samples. Moreover, these exosomes can be used to further diagnosis and therapeutics in veterinary pre-clinical and clinical studies.

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Exosomes isolation and identification from equine mesenchymal stem cells

BMC Veterinary Research ◽

10.1186/s12917-019-1789-9 ◽

2019 ◽

Vol 15 (1) ◽

Cited By ~ 13

Author(s):

Michele Christian Klymiuk ◽

Natalie Balz ◽

Mohamed I. Elashry ◽

Manuela Heimann ◽

Sabine Wenisch ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Isolation And Identification

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Effect of different culture conditions on nestin expression in fetal pancreas derived mesenchymal stem cells

Cytotherapy ◽

10.1016/j.jcyt.2017.02.222 ◽

2017 ◽

Vol 19 (5) ◽

pp. S148-S149

Author(s):

H. Aghayan ◽

k Falahzadeh ◽

B. Arjmand ◽

B. Larijani

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Culture Conditions ◽

Nestin Expression ◽

Fetal Pancreas

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STEM-19. THE ISOLATION AND IDENTIFICATION OF GLIOMA STEM CELLS AND MESENCHYMAL STEM CELLS FROM HUMAN GLIOMA SPECIMENS

Neuro-Oncology ◽

10.1093/neuonc/noaa215.836 ◽

2020 ◽

Vol 22 (Supplement_2) ◽

pp. ii200-ii200

Author(s):

Yan-jiao Yu ◽

Wang Jing ◽

Zhong-ping Chen

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Tumor Microenvironment ◽

Cell Suspension ◽

Primary Culture ◽

Glioma Stem Cells ◽

Isolation And Identification ◽

Primary Malignancy ◽

The Relationship

Abstract INTRODUCTION Glioma is the primary malignancy with the highest incidence in central nervous system. Studies have shown that glioma stem cell (GSC) is the important origin for recurrence and closely associated with tumor microenvironment. Glioma-associated human mesenchymal stem cells (GA-hMSCs), as an important part of the tumor microenvironment, might promote proliferation and invasion of GSC. We try to isolate GSCs and GA-hMSCs through glioma primary culture, and investigate the relationship between GSC and GA-hMSCs, as well as their roles in the recurrences of glioma. MATERIALS AND METHODS Five surgical specimens from patients with human high-grade glioma were collected for primary culture. The surgical specimens were cut up and digested with Dispase II (collagenase), terminated by adding plenty of DNase II. Cell suspension was collected and resuspended in conditional medium for MSC or GSC. After being cultured for several passages, GSCs were identified by immunofluorescence, and GA-hMSCs were identified by flow cytometry and in vitro differentiation assays. RESULTS Cell cultures were observed with an inverted phase-contrast microscope after 24 hours. Adherent GA-hMSCs were collected and the supernatant was discarded with non-adherent cells. GSC was cultured as neurospheres. The single-cell suspension was prepared from spheres of GSC to identify their surface expression profile by immunofluorescence assays after several passages. The results confirmed that GSCs positively expressed CD133, Nestin and SOX2, but did not express GFAP and Tublin III. GA-hMSCs positively expressed CD105, CD73, CD90 and negatively expressed CD45 and CD34. GA-hMSCs could differentiate into adipocytes, chondrocytes and osteocytes. CONCLUSION We isolated and identified GSCs and GA-hMSCs from glioma primary culture. Our future work will focus on the relationship between the two types of cells and their roles in the process of tumor recurrence based on the successful isolation of GSC and GA-hMSCs. Keywords: Glioma; Mesenchymal stem cells; Glioma stem cells; Recurrence

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Isolation and identification of exosomes from feline plasma, urine and adipose-derived mesenchymal stem cells

BMC Veterinary Research ◽

10.1186/s12917-021-02960-4 ◽

2021 ◽

Vol 17 (1) ◽

Author(s):

Dongsheng Li ◽

Huina Luo ◽

Huimin Ruan ◽

Zhisheng Chen ◽

Shengfeng Chen ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Transmembrane Protein ◽

Cell Communication ◽

Phospholipid Bilayer ◽

Differential Centrifugation ◽

Isolation And Identification ◽

Feasible Method ◽

Transmission Electron ◽

Mediate Cell

Abstract Background Exosomes, internal proteins, lipids, and nucleic acids coated by phospholipid bilayer membranes, are one type of small extracellular vesicles, which can mediate cell-cell communication. In recent years, exosomes have gained considerable scientific interest due to their widely applied prospect in the diagnosis and therapeutics of human and animal diseases. In this study, we describe for the first time a feasible method designed to isolate and characterize exosomes from feline plasma, urine and adipose-derived mesenchymal stem cells. Results Exosomes from feline plasma, urine and adipose-derived mesenchymal stem cells were successfully isolated by differential centrifugation. Quantification and sizing of exosomes were assessed by transmission electron microscopy, flow nano analysis and western blotting. Detected particles showed the normal size (30–100 nm) and morphology described for exosomes, as well as presence of the transmembrane protein (TSG101, CD9, CD63, and CD81) known as exosomal marker. Conclusions The results suggest that differential centrifugation is a feasible method for isolation of exosomes from different types of feline samples. Moreover, these exosomes can be used to further diagnosis and therapeutics in veterinary pre-clinical and clinical studies.

Download Full-text