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Extracellular Matrix and Growth Factors Improve the Efficacy of Intramuscular Islet Transplantation.

Tsuchiya H, Sakata N, Yoshimatsu G, Fukase M, Aoki T, Ishida M, Katayose Y, Egawa S, Unno M - PLoS ONE (2015)

Bottom Line: The following three groups were evaluated: islets without treatment (islets-only group), islets embedded in ECM with growth factors (Matrigel group), and islets embedded in ECM without growth factors [growth factor-reduced (GFR) Matrigel group].The viability and insulin-releasing function of islets cultured for 96 h were significantly improved in Matrigel and GFR Matrigel groups compared with the islets-only group.On histological examination, significantly decreased numbers of TdT-mediated deoxyuridine triphosphate-biotin nick end labeling-positive islet cells and significantly increased numbers of Ki67-positive cells were observed in the Matrigel and GFR Matrigel groups at POD 3.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Tohoku University, Sendai, Japan.

ABSTRACT

Background: The efficacy of intramuscular islet transplantation is poor despite being technically simple, safe, and associated with reduced rates of severe complications. We evaluated the efficacy of combined treatment with extracellular matrix (ECM) and growth factors in intramuscular islet transplantation.

Methods: Male BALB/C mice were used for the in vitro and transplantation studies. The following three groups were evaluated: islets without treatment (islets-only group), islets embedded in ECM with growth factors (Matrigel group), and islets embedded in ECM without growth factors [growth factor-reduced (GFR) Matrigel group]. The viability and insulin-releasing function of islets cultured for 96 h were significantly improved in Matrigel and GFR Matrigel groups compared with the islets-only group.

Results: Blood glucose and serum insulin levels immediately following transplantation were significantly improved in the Matrigel and GFR Matrigel groups and remained significantly improved in the Matrigel group at postoperative day (POD) 28. On histological examination, significantly decreased numbers of TdT-mediated deoxyuridine triphosphate-biotin nick end labeling-positive islet cells and significantly increased numbers of Ki67-positive cells were observed in the Matrigel and GFR Matrigel groups at POD 3. Peri-islet revascularization was most prominent in the Matrigel group at POD 14.

Conclusions: The efficacy of intramuscular islet transplantation was improved by combination treatment with ECM and growth factors through the inhibition of apoptosis, increased proliferation of islet cells, and promotion of revascularization.

No MeSH data available.


Related in: MedlinePlus

Intracellular signaling activity in islets cultured in Matrigel.FAK, Erk, and Akt protein levels in islets were evaluated by Western immunoblotting to assess the effect of Matrigel on cellular growth and cytoprotection. Cell lysates obtained from the islets-only, Matrigel, and GFR Matrigel groups were subjected to SDS-PAGE and Western immunoblotting using the indicated antibodies. Glyceraldehyde, phosphate dehydrogenase was used as the loading control. Phosphorylated FAK, Erk, and Akt were strongly expressed in the Matrigel and GFR Matrigel groups compared with the islets-only group. Erk, extracellular signal-regulated kinase; FAK, focal adhesion kinase; GFR, growth factor-reduced; SDS, sodium dodecyl sulfate.
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pone.0140910.g003: Intracellular signaling activity in islets cultured in Matrigel.FAK, Erk, and Akt protein levels in islets were evaluated by Western immunoblotting to assess the effect of Matrigel on cellular growth and cytoprotection. Cell lysates obtained from the islets-only, Matrigel, and GFR Matrigel groups were subjected to SDS-PAGE and Western immunoblotting using the indicated antibodies. Glyceraldehyde, phosphate dehydrogenase was used as the loading control. Phosphorylated FAK, Erk, and Akt were strongly expressed in the Matrigel and GFR Matrigel groups compared with the islets-only group. Erk, extracellular signal-regulated kinase; FAK, focal adhesion kinase; GFR, growth factor-reduced; SDS, sodium dodecyl sulfate.

Mentions: Regarding intracellular signaling activity in islets cultured in ECM, increased expression of phosphorylated FAK, Erk, and Akt was clearly demonstrated in the Matrigel and GFR Matrigel groups compared with the islets-only group (Fig 3). These data indicate culture in ECM stimulates MAPK/Erk and PI3K/Akt pathways, contributing to the promotion of islet cellular proliferation and survival.


Extracellular Matrix and Growth Factors Improve the Efficacy of Intramuscular Islet Transplantation.

Tsuchiya H, Sakata N, Yoshimatsu G, Fukase M, Aoki T, Ishida M, Katayose Y, Egawa S, Unno M - PLoS ONE (2015)

Intracellular signaling activity in islets cultured in Matrigel.FAK, Erk, and Akt protein levels in islets were evaluated by Western immunoblotting to assess the effect of Matrigel on cellular growth and cytoprotection. Cell lysates obtained from the islets-only, Matrigel, and GFR Matrigel groups were subjected to SDS-PAGE and Western immunoblotting using the indicated antibodies. Glyceraldehyde, phosphate dehydrogenase was used as the loading control. Phosphorylated FAK, Erk, and Akt were strongly expressed in the Matrigel and GFR Matrigel groups compared with the islets-only group. Erk, extracellular signal-regulated kinase; FAK, focal adhesion kinase; GFR, growth factor-reduced; SDS, sodium dodecyl sulfate.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4608691&req=5

pone.0140910.g003: Intracellular signaling activity in islets cultured in Matrigel.FAK, Erk, and Akt protein levels in islets were evaluated by Western immunoblotting to assess the effect of Matrigel on cellular growth and cytoprotection. Cell lysates obtained from the islets-only, Matrigel, and GFR Matrigel groups were subjected to SDS-PAGE and Western immunoblotting using the indicated antibodies. Glyceraldehyde, phosphate dehydrogenase was used as the loading control. Phosphorylated FAK, Erk, and Akt were strongly expressed in the Matrigel and GFR Matrigel groups compared with the islets-only group. Erk, extracellular signal-regulated kinase; FAK, focal adhesion kinase; GFR, growth factor-reduced; SDS, sodium dodecyl sulfate.
Mentions: Regarding intracellular signaling activity in islets cultured in ECM, increased expression of phosphorylated FAK, Erk, and Akt was clearly demonstrated in the Matrigel and GFR Matrigel groups compared with the islets-only group (Fig 3). These data indicate culture in ECM stimulates MAPK/Erk and PI3K/Akt pathways, contributing to the promotion of islet cellular proliferation and survival.

Bottom Line: The following three groups were evaluated: islets without treatment (islets-only group), islets embedded in ECM with growth factors (Matrigel group), and islets embedded in ECM without growth factors [growth factor-reduced (GFR) Matrigel group].The viability and insulin-releasing function of islets cultured for 96 h were significantly improved in Matrigel and GFR Matrigel groups compared with the islets-only group.On histological examination, significantly decreased numbers of TdT-mediated deoxyuridine triphosphate-biotin nick end labeling-positive islet cells and significantly increased numbers of Ki67-positive cells were observed in the Matrigel and GFR Matrigel groups at POD 3.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Tohoku University, Sendai, Japan.

ABSTRACT

Background: The efficacy of intramuscular islet transplantation is poor despite being technically simple, safe, and associated with reduced rates of severe complications. We evaluated the efficacy of combined treatment with extracellular matrix (ECM) and growth factors in intramuscular islet transplantation.

Methods: Male BALB/C mice were used for the in vitro and transplantation studies. The following three groups were evaluated: islets without treatment (islets-only group), islets embedded in ECM with growth factors (Matrigel group), and islets embedded in ECM without growth factors [growth factor-reduced (GFR) Matrigel group]. The viability and insulin-releasing function of islets cultured for 96 h were significantly improved in Matrigel and GFR Matrigel groups compared with the islets-only group.

Results: Blood glucose and serum insulin levels immediately following transplantation were significantly improved in the Matrigel and GFR Matrigel groups and remained significantly improved in the Matrigel group at postoperative day (POD) 28. On histological examination, significantly decreased numbers of TdT-mediated deoxyuridine triphosphate-biotin nick end labeling-positive islet cells and significantly increased numbers of Ki67-positive cells were observed in the Matrigel and GFR Matrigel groups at POD 3. Peri-islet revascularization was most prominent in the Matrigel group at POD 14.

Conclusions: The efficacy of intramuscular islet transplantation was improved by combination treatment with ECM and growth factors through the inhibition of apoptosis, increased proliferation of islet cells, and promotion of revascularization.

No MeSH data available.


Related in: MedlinePlus