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Activated pancreatic stellate cells can impair pancreatic islet function in mice.

Zang G, Sandberg M, Carlsson PO, Welsh N, Jansson L, Barbu A - Ups. J. Med. Sci. (2015)

Bottom Line: Furthermore, we measured their cytokine production and determined their effects on insulin release from simultaneously cultured islets.In islet transplants they were found mainly in the graft periphery.Increased islet cell caspase-3 activation and a decreased islet cell replication were consistently observed after co-culture for 2 or 7 days.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Cell Biology, Uppsala University , Uppsala , Sweden.

ABSTRACT

Background: Pancreatic or islet fibrosis is often associated with activated pancreatic stellate cells (PSCs). PSCs are considered not only to promote fibrosis, but also to be associated with glucose intolerance in some diseases. We therefore evaluated morphological and functional relationships between islets and PSCs in the normal mouse pancreas and transplanted islets.

Methods: Immunohistochemistry was used to map the presence of PSCs in the normal mouse pancreas and islets implanted under the renal capsule. We isolated and cultured mouse PSCs and characterized them morphologically by immunofluorescence staining. Furthermore, we measured their cytokine production and determined their effects on insulin release from simultaneously cultured islets.

Results: PSCs were scattered throughout the pancreas, with occasional cells within the islets, particularly in the islet capsule. In islet transplants they were found mainly in the graft periphery. Cultured PSCs became functionally activated and produced several cytokines. Throughout the culture period they linearly increased their production of interleukin-6 and mammalian keratinocyte-derived chemokine. PSC cytokine production was not affected by acute hyperglycemia. Syngeneic islets co-cultured with PSCs for 24-48 h increased their insulin release and lowered their insulin content. However, short-term insulin release in batch-type incubations was unaffected after 48 h of co-culture. Increased islet cell caspase-3 activation and a decreased islet cell replication were consistently observed after co-culture for 2 or 7 days.

Conclusion: Activated PSCs may contribute to impaired islet endocrine function seen in exocrine pancreatitis and in islet fibrosis associated with some cases of type 2 diabetes.

No MeSH data available.


Related in: MedlinePlus

Isolation and identification of mouse pancreatic stellate cells. A: Pancreatic stellate cell from mouse cultured for 24 h. There are numerous lipid droplets in the cytoplasm. B: Activated pancreatic stellate cells have lost lipid droplets and attained a fibroblastoid appearance. C–E: Isolated PSCs were cultured on culture slides, and their protein expression was analyzed as described in ‘Materials and methods’. C: Desmin (green) and Nile red (denoting lipid droplet presence) (red) fluorescence in PSCs cultured for 2 days after isolation. D: Desmin (green) and α-SMA (red) immunoreactivities in PSCs cultured for 14 days after isolation. 20× magnification. E: Vimentin (green) and -SMA (red) immunoreactivities in PSCs for 14 days after isolation. Scale bars 50 m (A and C) and 100 m (B, D, E). In all micrographs Hoechst 33258 was used for nuclear staining (blue).
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Figure 2: Isolation and identification of mouse pancreatic stellate cells. A: Pancreatic stellate cell from mouse cultured for 24 h. There are numerous lipid droplets in the cytoplasm. B: Activated pancreatic stellate cells have lost lipid droplets and attained a fibroblastoid appearance. C–E: Isolated PSCs were cultured on culture slides, and their protein expression was analyzed as described in ‘Materials and methods’. C: Desmin (green) and Nile red (denoting lipid droplet presence) (red) fluorescence in PSCs cultured for 2 days after isolation. D: Desmin (green) and α-SMA (red) immunoreactivities in PSCs cultured for 14 days after isolation. 20× magnification. E: Vimentin (green) and -SMA (red) immunoreactivities in PSCs for 14 days after isolation. Scale bars 50 m (A and C) and 100 m (B, D, E). In all micrographs Hoechst 33258 was used for nuclear staining (blue).

Mentions: We could routinely isolate and culture PSCs with a purity of more than 95%. Immediately after isolation they contained cytoplasmic lipid droplets (Figure 2A). After culture for a few days the PSCs became activated, as represented by morphological changes with disappearance of lipid droplets and a more fibroblastoid appearance (Figure 2B). Moreover, PSCs were immuno-positive for desmin (Figure 2C, D), vimentin (Figure 2E), and α-SMA (Figure 2D, E). Up to 3 days after isolation, desmin expression was doubled by the peri-nuclear presence of lipid droplets in the cytoplasm, as assessed by Nile red staining (Figure 2C). At later stages (10–14 days of culturing) desmin and vimentin expression was paralleled by a strong -SMA immunoreactivity (Figure 2D, E).


Activated pancreatic stellate cells can impair pancreatic islet function in mice.

Zang G, Sandberg M, Carlsson PO, Welsh N, Jansson L, Barbu A - Ups. J. Med. Sci. (2015)

Isolation and identification of mouse pancreatic stellate cells. A: Pancreatic stellate cell from mouse cultured for 24 h. There are numerous lipid droplets in the cytoplasm. B: Activated pancreatic stellate cells have lost lipid droplets and attained a fibroblastoid appearance. C–E: Isolated PSCs were cultured on culture slides, and their protein expression was analyzed as described in ‘Materials and methods’. C: Desmin (green) and Nile red (denoting lipid droplet presence) (red) fluorescence in PSCs cultured for 2 days after isolation. D: Desmin (green) and α-SMA (red) immunoreactivities in PSCs cultured for 14 days after isolation. 20× magnification. E: Vimentin (green) and -SMA (red) immunoreactivities in PSCs for 14 days after isolation. Scale bars 50 m (A and C) and 100 m (B, D, E). In all micrographs Hoechst 33258 was used for nuclear staining (blue).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4526872&req=5

Figure 2: Isolation and identification of mouse pancreatic stellate cells. A: Pancreatic stellate cell from mouse cultured for 24 h. There are numerous lipid droplets in the cytoplasm. B: Activated pancreatic stellate cells have lost lipid droplets and attained a fibroblastoid appearance. C–E: Isolated PSCs were cultured on culture slides, and their protein expression was analyzed as described in ‘Materials and methods’. C: Desmin (green) and Nile red (denoting lipid droplet presence) (red) fluorescence in PSCs cultured for 2 days after isolation. D: Desmin (green) and α-SMA (red) immunoreactivities in PSCs cultured for 14 days after isolation. 20× magnification. E: Vimentin (green) and -SMA (red) immunoreactivities in PSCs for 14 days after isolation. Scale bars 50 m (A and C) and 100 m (B, D, E). In all micrographs Hoechst 33258 was used for nuclear staining (blue).
Mentions: We could routinely isolate and culture PSCs with a purity of more than 95%. Immediately after isolation they contained cytoplasmic lipid droplets (Figure 2A). After culture for a few days the PSCs became activated, as represented by morphological changes with disappearance of lipid droplets and a more fibroblastoid appearance (Figure 2B). Moreover, PSCs were immuno-positive for desmin (Figure 2C, D), vimentin (Figure 2E), and α-SMA (Figure 2D, E). Up to 3 days after isolation, desmin expression was doubled by the peri-nuclear presence of lipid droplets in the cytoplasm, as assessed by Nile red staining (Figure 2C). At later stages (10–14 days of culturing) desmin and vimentin expression was paralleled by a strong -SMA immunoreactivity (Figure 2D, E).

Bottom Line: Furthermore, we measured their cytokine production and determined their effects on insulin release from simultaneously cultured islets.In islet transplants they were found mainly in the graft periphery.Increased islet cell caspase-3 activation and a decreased islet cell replication were consistently observed after co-culture for 2 or 7 days.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Cell Biology, Uppsala University , Uppsala , Sweden.

ABSTRACT

Background: Pancreatic or islet fibrosis is often associated with activated pancreatic stellate cells (PSCs). PSCs are considered not only to promote fibrosis, but also to be associated with glucose intolerance in some diseases. We therefore evaluated morphological and functional relationships between islets and PSCs in the normal mouse pancreas and transplanted islets.

Methods: Immunohistochemistry was used to map the presence of PSCs in the normal mouse pancreas and islets implanted under the renal capsule. We isolated and cultured mouse PSCs and characterized them morphologically by immunofluorescence staining. Furthermore, we measured their cytokine production and determined their effects on insulin release from simultaneously cultured islets.

Results: PSCs were scattered throughout the pancreas, with occasional cells within the islets, particularly in the islet capsule. In islet transplants they were found mainly in the graft periphery. Cultured PSCs became functionally activated and produced several cytokines. Throughout the culture period they linearly increased their production of interleukin-6 and mammalian keratinocyte-derived chemokine. PSC cytokine production was not affected by acute hyperglycemia. Syngeneic islets co-cultured with PSCs for 24-48 h increased their insulin release and lowered their insulin content. However, short-term insulin release in batch-type incubations was unaffected after 48 h of co-culture. Increased islet cell caspase-3 activation and a decreased islet cell replication were consistently observed after co-culture for 2 or 7 days.

Conclusion: Activated PSCs may contribute to impaired islet endocrine function seen in exocrine pancreatitis and in islet fibrosis associated with some cases of type 2 diabetes.

No MeSH data available.


Related in: MedlinePlus