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Effect of mesenchymal stem cells transplantation on glycaemic profile & their localization in streptozotocin induced diabetic Wistar rats.

Bhansali S, Kumar V, Saikia UN, Medhi B, Jha V, Bhansali A, Dutta P - Indian J. Med. Res. (2015)

Bottom Line: Administration of mesenchymal stem cells (MSCs) in irradiated diabetic rat model has transiently shown to decrease blood glucose level.There was a significant reduction in blood glucose level after administration of MSCs in the experimental group (P<0.001).Co-expression of anti-BrdU and anti-insulin antibody indicated trans-differentiation / fusion into insulin producing cells evidenced by significant increase in total number of islet (P=0.004) and insulin positive cells ( P<0.0001) in experimental group.

View Article: PubMed Central - PubMed

Affiliation: Department of Endocrinology, Postgraduate Institute of Medical Education & Research, Chandigarh, India.

ABSTRACT

Background & objectives: Bone marrow is a rich source of adult stem cells that can differentiate into various cell types. Administration of mesenchymal stem cells (MSCs) in irradiated diabetic rat model has transiently shown to decrease blood glucose level. This study examines the effect of high dose and multiple injections of MSCs on glycemic profile, their localization and regeneration of islet in diabetic Wistar rat.

Methods: The study was carried out in male Wistar rats categorized into three groups (n=6, in each group): Group 1 as control, group 2 streptozotocin (STZ) (50 mg/kg) induced diabetic group and group 3 experimental group; 5-bromo-2-deoxyuridine (BrdU) labelled allogenic MSCs were injected in the non-irradiated diabetic rat of the experimental group through tail vein. The blood glucose profile was subsequently monitored at regular intervals. Rats were sacrificed on day 45 and pancreas was examined for localization of BrdU labelled stem cells by immunofluorescence and islet-neogenesis by immunohistochemistry .

Results: There was a significant reduction in blood glucose level after administration of MSCs in the experimental group (P<0.001). The presence of BrdU labelled MSCs in islet suggested their localization in the pancreas. Co-expression of anti-BrdU and anti-insulin antibody indicated trans-differentiation / fusion into insulin producing cells evidenced by significant increase in total number of islet (P=0.004) and insulin positive cells ( P<0.0001) in experimental group.

Interpretation & conclusions: Our results showed that the MSCs administration in non-irradiated diabetic Wistar rat reduced hyperglycaemia and was accompanied by increased islet-neogenesis, possibly through trans-differentiation/fusion.

No MeSH data available.


Related in: MedlinePlus

(A). Photomicrographs of cultured mesenchymal stem cells (20X magnification), (B) Flow cytometry analysis of percentage of dual positive (CD29 and CD54) cultured mesenchymal stem cells, (C, D) Flow cytometry analysis of percentage of negative marker (CD 34 and CD 45) for haematopoietic lineage, and (E) Flow cytometry analysis of percentage of labelling of mesenchymal stem cells with anti- BrdU.
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Figure 1: (A). Photomicrographs of cultured mesenchymal stem cells (20X magnification), (B) Flow cytometry analysis of percentage of dual positive (CD29 and CD54) cultured mesenchymal stem cells, (C, D) Flow cytometry analysis of percentage of negative marker (CD 34 and CD 45) for haematopoietic lineage, and (E) Flow cytometry analysis of percentage of labelling of mesenchymal stem cells with anti- BrdU.

Mentions: Isolation, characterisation and labelling of MSCs: After in vitro culture for 10-12 days cells having fibroblast like morphology adhered to flask surface (Fig. 1A). Flow cytometeric analysis showed a high expression of CD29 and CD54 (Fig. 1B) and negative staining for CD34 and CD45 (Fig. 1C, D) markers indicating that cultured cells were of mesenchymal origin as well as of high purity without haematopoietic lineage cell contamination. Before transplantation, the nuclei of MSCs were labelled with BrdU. Further, the efficacy of labelling was assessed by staining with anti-BrdU antibody and flow cytometery revealed 90 per cent (n=6) of these MSCs were stained positively for BrdU (Fig. 1E) and cell viability (92%) with trypan blue.


Effect of mesenchymal stem cells transplantation on glycaemic profile & their localization in streptozotocin induced diabetic Wistar rats.

Bhansali S, Kumar V, Saikia UN, Medhi B, Jha V, Bhansali A, Dutta P - Indian J. Med. Res. (2015)

(A). Photomicrographs of cultured mesenchymal stem cells (20X magnification), (B) Flow cytometry analysis of percentage of dual positive (CD29 and CD54) cultured mesenchymal stem cells, (C, D) Flow cytometry analysis of percentage of negative marker (CD 34 and CD 45) for haematopoietic lineage, and (E) Flow cytometry analysis of percentage of labelling of mesenchymal stem cells with anti- BrdU.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4557252&req=5

Figure 1: (A). Photomicrographs of cultured mesenchymal stem cells (20X magnification), (B) Flow cytometry analysis of percentage of dual positive (CD29 and CD54) cultured mesenchymal stem cells, (C, D) Flow cytometry analysis of percentage of negative marker (CD 34 and CD 45) for haematopoietic lineage, and (E) Flow cytometry analysis of percentage of labelling of mesenchymal stem cells with anti- BrdU.
Mentions: Isolation, characterisation and labelling of MSCs: After in vitro culture for 10-12 days cells having fibroblast like morphology adhered to flask surface (Fig. 1A). Flow cytometeric analysis showed a high expression of CD29 and CD54 (Fig. 1B) and negative staining for CD34 and CD45 (Fig. 1C, D) markers indicating that cultured cells were of mesenchymal origin as well as of high purity without haematopoietic lineage cell contamination. Before transplantation, the nuclei of MSCs were labelled with BrdU. Further, the efficacy of labelling was assessed by staining with anti-BrdU antibody and flow cytometery revealed 90 per cent (n=6) of these MSCs were stained positively for BrdU (Fig. 1E) and cell viability (92%) with trypan blue.

Bottom Line: Administration of mesenchymal stem cells (MSCs) in irradiated diabetic rat model has transiently shown to decrease blood glucose level.There was a significant reduction in blood glucose level after administration of MSCs in the experimental group (P<0.001).Co-expression of anti-BrdU and anti-insulin antibody indicated trans-differentiation / fusion into insulin producing cells evidenced by significant increase in total number of islet (P=0.004) and insulin positive cells ( P<0.0001) in experimental group.

View Article: PubMed Central - PubMed

Affiliation: Department of Endocrinology, Postgraduate Institute of Medical Education & Research, Chandigarh, India.

ABSTRACT

Background & objectives: Bone marrow is a rich source of adult stem cells that can differentiate into various cell types. Administration of mesenchymal stem cells (MSCs) in irradiated diabetic rat model has transiently shown to decrease blood glucose level. This study examines the effect of high dose and multiple injections of MSCs on glycemic profile, their localization and regeneration of islet in diabetic Wistar rat.

Methods: The study was carried out in male Wistar rats categorized into three groups (n=6, in each group): Group 1 as control, group 2 streptozotocin (STZ) (50 mg/kg) induced diabetic group and group 3 experimental group; 5-bromo-2-deoxyuridine (BrdU) labelled allogenic MSCs were injected in the non-irradiated diabetic rat of the experimental group through tail vein. The blood glucose profile was subsequently monitored at regular intervals. Rats were sacrificed on day 45 and pancreas was examined for localization of BrdU labelled stem cells by immunofluorescence and islet-neogenesis by immunohistochemistry .

Results: There was a significant reduction in blood glucose level after administration of MSCs in the experimental group (P<0.001). The presence of BrdU labelled MSCs in islet suggested their localization in the pancreas. Co-expression of anti-BrdU and anti-insulin antibody indicated trans-differentiation / fusion into insulin producing cells evidenced by significant increase in total number of islet (P=0.004) and insulin positive cells ( P<0.0001) in experimental group.

Interpretation & conclusions: Our results showed that the MSCs administration in non-irradiated diabetic Wistar rat reduced hyperglycaemia and was accompanied by increased islet-neogenesis, possibly through trans-differentiation/fusion.

No MeSH data available.


Related in: MedlinePlus