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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

(A-D): undifferentiated VSELs grown on MEF cells, colony formation of VSELs after 3 days, Bar = 100 µm. A) A colony of VSEL stem cells (40×); B) A colony of VSEL stem cells (100×), round bright cells are VSELs; C) a colony of VSEL stem cells (200×); D) A colony of VSEL stem cells (400×) (image was taken by Olympus IX71 inverted microscope). (Image was taken by Olympus BX51 microscope).
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Figure 0001: (A-D): undifferentiated VSELs grown on MEF cells, colony formation of VSELs after 3 days, Bar = 100 µm. A) A colony of VSEL stem cells (40×); B) A colony of VSEL stem cells (100×), round bright cells are VSELs; C) a colony of VSEL stem cells (200×); D) A colony of VSEL stem cells (400×) (image was taken by Olympus IX71 inverted microscope). (Image was taken by Olympus BX51 microscope).

Mentions: For best results, VSEL stem cells were cultivated by two different ways. One group was proliferated on a feeder layer of mitotically inactivated MEF in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% FBS, L-glutamine (2 mM, Gibco), ß-mercaptoethanol (final concentration 5×105M), penicillin/ streptomycin (100 U/ml penicillin and 100 µg/ml streptomycin (Gibco/BRL) and non essential amino acid (NEAA, stock solution diluted 1:100, Gibco). MEF cells secrete special chemical mediators that affect VSEL stem cells in a paracrine manner and inhibit differentiation. Another group was proliferated on coated dishes with the same culture media, plus LIF (10 ng/ ml) (Sigma, L5158) (11), and the dishes were coated with 0/1% gelatin solution (Figure 1).


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)

(A-D): undifferentiated VSELs grown on MEF cells, colony formation of VSELs after 3 days, Bar = 100 µm. A) A colony of VSEL stem cells (40×); B) A colony of VSEL stem cells (100×), round bright cells are VSELs; C) a colony of VSEL stem cells (200×); D) A colony of VSEL stem cells (400×) (image was taken by Olympus IX71 inverted microscope). (Image was taken by Olympus BX51 microscope).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 0001: (A-D): undifferentiated VSELs grown on MEF cells, colony formation of VSELs after 3 days, Bar = 100 µm. A) A colony of VSEL stem cells (40×); B) A colony of VSEL stem cells (100×), round bright cells are VSELs; C) a colony of VSEL stem cells (200×); D) A colony of VSEL stem cells (400×) (image was taken by Olympus IX71 inverted microscope). (Image was taken by Olympus BX51 microscope).
Mentions: For best results, VSEL stem cells were cultivated by two different ways. One group was proliferated on a feeder layer of mitotically inactivated MEF in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% FBS, L-glutamine (2 mM, Gibco), ß-mercaptoethanol (final concentration 5×105M), penicillin/ streptomycin (100 U/ml penicillin and 100 µg/ml streptomycin (Gibco/BRL) and non essential amino acid (NEAA, stock solution diluted 1:100, Gibco). MEF cells secrete special chemical mediators that affect VSEL stem cells in a paracrine manner and inhibit differentiation. Another group was proliferated on coated dishes with the same culture media, plus LIF (10 ng/ ml) (Sigma, L5158) (11), and the dishes were coated with 0/1% gelatin solution (Figure 1).

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