Limits...
Intravenous transplantation of very small embryonic like stem cells in treatment of diabetes mellitus.

Abouzaripour M, Ragerdi Kashani I, Pasbakhsh P, Atlasy N - Avicenna J Med Biotechnol (2015 Jan-Mar)

Bottom Line: It was proved that CD45-, CXCR4+, and Sca1+ sorted cells express oct4 and SSEA1.Our results revealed that intravenously implanted VSELs could migrate into the pancreas of hosts and survive in the diabetic pancreas.In treated groups, blood glucose decreased significantly for at least two month and the weights of mice increased gradually.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran.

ABSTRACT

Background: Diabetes Mellitus (DM), simply known as diabetes, refers to a group of metabolic diseases in which there are high blood sugar levels over a prolonged period. In this study, the feasibility and safety of intravenous transplantation of Very Small Embryonic Like stem cells (VSELs) were investigated for diabetes repair, and finally the migration and distribution of these cells in hosts were observed.

Methods: Mouse bone marrow VSELs were isolated by Fluorescent Activating Cell Sorting (FACS) method by using fluorescent antibodies against CD45, CXCR4 and Sca1 markers. Sorted cells were analyzed for expression of oct4 and SSEA1 markers with immunocytochemistry staining method. To determine multilineage differentiation, sorted cells were differentiated to Schwann, osteocyte and beta cells. Ten days after the establishment of a mouse model of pancreas necrosis, DiI-labeled VSELs were injected into these mice via tail vein. Pancreases were harvested 4 weeks after transplantation and the sections of these tissues were observed under fluorescent microscope.

Results: It was proved that CD45-, CXCR4+, and Sca1+ sorted cells express oct4 and SSEA1. Our results revealed that intravenously implanted VSELs could migrate into the pancreas of hosts and survive in the diabetic pancreas. In treated groups, blood glucose decreased significantly for at least two month and the weights of mice increased gradually.

Conclusion: This study provides a strategy for using VSELs for curing diabetes and other regenerative diseases, and the strategy is considered an alternative for other stem cell types.

No MeSH data available.


Related in: MedlinePlus

Immunofluorescence staining of VSEL stem cell. CD45−, Sca1+, and CXCR4+ cells are SSEA1+ and express oct4; A) Oct4 (green) expression; B) SSEA1 (red) expression. Nuclei visualized after DAPI staining (40×) (image was taken by Olympus IX71 inverted microscope).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4388887&req=5

Figure 0002: Immunofluorescence staining of VSEL stem cell. CD45−, Sca1+, and CXCR4+ cells are SSEA1+ and express oct4; A) Oct4 (green) expression; B) SSEA1 (red) expression. Nuclei visualized after DAPI staining (40×) (image was taken by Olympus IX71 inverted microscope).

Mentions: The expression of each antigen was examined in cells from two independent sorts. After passage 1, the CD45−/Sca1+ /CXCr4+ sorted cells were fixed for 2 hr at room temperature with 4% paraformaldehyde and then incubated at room temperature for 15 min with 1% Triton ×-100/phosphate-buffered saline (PBS). Cells were washed three times in PBS and blocked at 37°C for over 3 hr with 4% normal goat serum (Chemicon). Subsequently, cells were incubated at 4°C overnight with rabbit polyclonal primary antibody to oct4 (1:200, Abcam18976, USA), and mouse monoclonal primary antibody to SSEA1 (1:200, Abcam16285, USA). Cells were washed three times in PBS and incubated at 37°C for 2 hr with FITC goat anti rabbit (ab98511) and PE goat anti mouse secondary antibodies (1:500 in 1% normal goat serum in PBS). Unbound secondary antibodies were removed in three washes with PBS. Nuclei were identified by DAPI (Invitrogen) staining at a dilution of 1:10,000 at room temperature for 5 min. Images were acquired using an inverted microscope (Olympus, IX71, and Japan) (Figure 2).


Intravenous transplantation of very small embryonic like stem cells in treatment of diabetes mellitus.

Abouzaripour M, Ragerdi Kashani I, Pasbakhsh P, Atlasy N - Avicenna J Med Biotechnol (2015 Jan-Mar)

Immunofluorescence staining of VSEL stem cell. CD45−, Sca1+, and CXCR4+ cells are SSEA1+ and express oct4; A) Oct4 (green) expression; B) SSEA1 (red) expression. Nuclei visualized after DAPI staining (40×) (image was taken by Olympus IX71 inverted microscope).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4388887&req=5

Figure 0002: Immunofluorescence staining of VSEL stem cell. CD45−, Sca1+, and CXCR4+ cells are SSEA1+ and express oct4; A) Oct4 (green) expression; B) SSEA1 (red) expression. Nuclei visualized after DAPI staining (40×) (image was taken by Olympus IX71 inverted microscope).
Mentions: The expression of each antigen was examined in cells from two independent sorts. After passage 1, the CD45−/Sca1+ /CXCr4+ sorted cells were fixed for 2 hr at room temperature with 4% paraformaldehyde and then incubated at room temperature for 15 min with 1% Triton ×-100/phosphate-buffered saline (PBS). Cells were washed three times in PBS and blocked at 37°C for over 3 hr with 4% normal goat serum (Chemicon). Subsequently, cells were incubated at 4°C overnight with rabbit polyclonal primary antibody to oct4 (1:200, Abcam18976, USA), and mouse monoclonal primary antibody to SSEA1 (1:200, Abcam16285, USA). Cells were washed three times in PBS and incubated at 37°C for 2 hr with FITC goat anti rabbit (ab98511) and PE goat anti mouse secondary antibodies (1:500 in 1% normal goat serum in PBS). Unbound secondary antibodies were removed in three washes with PBS. Nuclei were identified by DAPI (Invitrogen) staining at a dilution of 1:10,000 at room temperature for 5 min. Images were acquired using an inverted microscope (Olympus, IX71, and Japan) (Figure 2).

Bottom Line: It was proved that CD45-, CXCR4+, and Sca1+ sorted cells express oct4 and SSEA1.Our results revealed that intravenously implanted VSELs could migrate into the pancreas of hosts and survive in the diabetic pancreas.In treated groups, blood glucose decreased significantly for at least two month and the weights of mice increased gradually.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran.

ABSTRACT

Background: Diabetes Mellitus (DM), simply known as diabetes, refers to a group of metabolic diseases in which there are high blood sugar levels over a prolonged period. In this study, the feasibility and safety of intravenous transplantation of Very Small Embryonic Like stem cells (VSELs) were investigated for diabetes repair, and finally the migration and distribution of these cells in hosts were observed.

Methods: Mouse bone marrow VSELs were isolated by Fluorescent Activating Cell Sorting (FACS) method by using fluorescent antibodies against CD45, CXCR4 and Sca1 markers. Sorted cells were analyzed for expression of oct4 and SSEA1 markers with immunocytochemistry staining method. To determine multilineage differentiation, sorted cells were differentiated to Schwann, osteocyte and beta cells. Ten days after the establishment of a mouse model of pancreas necrosis, DiI-labeled VSELs were injected into these mice via tail vein. Pancreases were harvested 4 weeks after transplantation and the sections of these tissues were observed under fluorescent microscope.

Results: It was proved that CD45-, CXCR4+, and Sca1+ sorted cells express oct4 and SSEA1. Our results revealed that intravenously implanted VSELs could migrate into the pancreas of hosts and survive in the diabetic pancreas. In treated groups, blood glucose decreased significantly for at least two month and the weights of mice increased gradually.

Conclusion: This study provides a strategy for using VSELs for curing diabetes and other regenerative diseases, and the strategy is considered an alternative for other stem cell types.

No MeSH data available.


Related in: MedlinePlus