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Allogeneic guinea pig mesenchymal stem cells ameliorate neurological changes in experimental colitis.

Stavely R, Robinson AM, Miller S, Boyd R, Sakkal S, Nurgali K - Stem Cell Res Ther (2015)

Bottom Line: This study aims to isolate and characterise guinea pig MSCs and then test their therapeutic potential for the treatment of enteric neuropathy associated with intestinal inflammation.MSCs from both sources secreted TGF-β1 which exerted neuroprotective effects in vitro.In vitro characteristics of MSCs cannot be extrapolated to their therapeutic efficacy.

View Article: PubMed Central - PubMed

Affiliation: Centre for Chronic Disease, College of Health and Biomedicine, Western Centre for Health, Research and Education, Sunshine Hospital, 176 Furlong road, Melbourne, 3021, Victoria, Australia. rhian.stavely@live.vu.edu.au.

ABSTRACT

Background: The use of mesenchymal stem cells (MSCs) to treat inflammatory bowel disease (IBD) is of great interest because of their immunomodulatory properties. Damage to the enteric nervous system (ENS) is implicated in IBD pathophysiology and disease progression. The most commonly used model to study inflammation-induced changes to the ENS is 2,4,6-trinitrobenzene-sulfonate acid (TNBS)-induced colitis in guinea pigs; however, no studies using guinea pig MSCs in colitis have been performed. This study aims to isolate and characterise guinea pig MSCs and then test their therapeutic potential for the treatment of enteric neuropathy associated with intestinal inflammation.

Methods: MSCs from guinea pig bone marrow and adipose tissue were isolated and characterised in vitro. In in vivo experiments, guinea pigs received either TNBS for the induction of colitis or sham treatment by enema. MSCs were administered at a dose of 1 × 10(6) cells via enema 3 h after the induction of colitis. Colon tissues were collected 24 and 72 h after TNBS administration to assess the level of inflammation and damage to the ENS. The secretion of transforming growth factor-β1 (TGF-β1) was analysed in MSC conditioned medium by flow cytometry.

Results: Cells isolated from both sources were adherent to plastic, multipotent and expressed some human MSC surface markers. In vitro characterisation revealed distinct differences in growth kinetics, clonogenicity and cell morphology between MSC types. In an in vivo model of TNBS-induced colitis, guinea pig bone marrow MSCs were comparatively more efficacious than adipose tissue MSCs in attenuating weight loss, colonic tissue damage and leukocyte infiltration into the mucosa and myenteric plexus. MSCs from both sources were equally neuroprotective in the amelioration of enteric neuronal loss and changes to the neurochemical coding of neuronal subpopulations. MSCs from both sources secreted TGF-β1 which exerted neuroprotective effects in vitro.

Conclusions: This study is the first evaluating the functional capacity of guinea pig bone marrow and adipose tissue-derived MSCs and providing evidence of their neuroprotective value in an animal model of colitis. In vitro characteristics of MSCs cannot be extrapolated to their therapeutic efficacy. TGF-β1 released by both types of MSCs might have contributed to the attenuation of enteric neuropathy associated with colitis.

No MeSH data available.


Related in: MedlinePlus

Immunophenotype and clonogenicity of guinea pig MSCs from bone marrow and adipose tissue. a GpBM-MSC and gpAT-MSCs were analysed for cell surface antigen expression of known positive MSC markers CD29 and CD73. Red closed histograms represent MSCs labelled with antibodies against the surface antigen indicated on the right-hand side of each row. Blue open histograms show isotype controls. GpBM-MSCs (b) and gpAT-MSCs (b′) adhered to plastic with a perceptible appearance typical of MSCs in culture. Scale bar = 200 μm. The clonogenicity of gpBM-MSCs (c) and gpAT-MSCs (c′) was determined by a colony-forming unit-fibroblast (CFU-f) assay (n = 4 independent cultures per group). d CFU-f counts were quantified as a percentage of the total viable cells seeded. ****P <0.0001. gpAT-MSC guinea pig adipose tissue-derived mesenchymal stem cell, gpBM-MSC guinea pig bone marrow-derived mesenchymal stem cell, MSC mesenchymal stem cell
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Fig1: Immunophenotype and clonogenicity of guinea pig MSCs from bone marrow and adipose tissue. a GpBM-MSC and gpAT-MSCs were analysed for cell surface antigen expression of known positive MSC markers CD29 and CD73. Red closed histograms represent MSCs labelled with antibodies against the surface antigen indicated on the right-hand side of each row. Blue open histograms show isotype controls. GpBM-MSCs (b) and gpAT-MSCs (b′) adhered to plastic with a perceptible appearance typical of MSCs in culture. Scale bar = 200 μm. The clonogenicity of gpBM-MSCs (c) and gpAT-MSCs (c′) was determined by a colony-forming unit-fibroblast (CFU-f) assay (n = 4 independent cultures per group). d CFU-f counts were quantified as a percentage of the total viable cells seeded. ****P <0.0001. gpAT-MSC guinea pig adipose tissue-derived mesenchymal stem cell, gpBM-MSC guinea pig bone marrow-derived mesenchymal stem cell, MSC mesenchymal stem cell

Mentions: To examine the immunophenotype of guinea pig MSCs, cells were analysed by flow cytometry for the presence of positive MSC markers CD29, CD44, CD73 and CD90 and negative MSC markers CD34 (hematopoietic progenitors) and CD45 (leukocyte common antigen). Positive expression of CD29 was observed in both gpBM-MSCs (95.1 %) and gpAT-MSCs (97.6 %). There was differential expression of the CD73 antigen such that 85.0 % of gpBM-MSCs expressed the antigen compared with negligible expression in gpAT-MSCs (0.2 %) (Fig. 1a). Negligible expression of the remaining surface markers was observed in both cell lines (gpBM-MSC: CD44, 1.9 %; CD90, 1.5 %; CD34, 4.4 %; CD45, 5.5 %. gpAT-MSC: CD44, 0.2 %; CD90, 0.2 %; CD34, 0.2 %; CD45, 0.7 %; data not shown).Fig. 1


Allogeneic guinea pig mesenchymal stem cells ameliorate neurological changes in experimental colitis.

Stavely R, Robinson AM, Miller S, Boyd R, Sakkal S, Nurgali K - Stem Cell Res Ther (2015)

Immunophenotype and clonogenicity of guinea pig MSCs from bone marrow and adipose tissue. a GpBM-MSC and gpAT-MSCs were analysed for cell surface antigen expression of known positive MSC markers CD29 and CD73. Red closed histograms represent MSCs labelled with antibodies against the surface antigen indicated on the right-hand side of each row. Blue open histograms show isotype controls. GpBM-MSCs (b) and gpAT-MSCs (b′) adhered to plastic with a perceptible appearance typical of MSCs in culture. Scale bar = 200 μm. The clonogenicity of gpBM-MSCs (c) and gpAT-MSCs (c′) was determined by a colony-forming unit-fibroblast (CFU-f) assay (n = 4 independent cultures per group). d CFU-f counts were quantified as a percentage of the total viable cells seeded. ****P <0.0001. gpAT-MSC guinea pig adipose tissue-derived mesenchymal stem cell, gpBM-MSC guinea pig bone marrow-derived mesenchymal stem cell, MSC mesenchymal stem cell
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4697327&req=5

Fig1: Immunophenotype and clonogenicity of guinea pig MSCs from bone marrow and adipose tissue. a GpBM-MSC and gpAT-MSCs were analysed for cell surface antigen expression of known positive MSC markers CD29 and CD73. Red closed histograms represent MSCs labelled with antibodies against the surface antigen indicated on the right-hand side of each row. Blue open histograms show isotype controls. GpBM-MSCs (b) and gpAT-MSCs (b′) adhered to plastic with a perceptible appearance typical of MSCs in culture. Scale bar = 200 μm. The clonogenicity of gpBM-MSCs (c) and gpAT-MSCs (c′) was determined by a colony-forming unit-fibroblast (CFU-f) assay (n = 4 independent cultures per group). d CFU-f counts were quantified as a percentage of the total viable cells seeded. ****P <0.0001. gpAT-MSC guinea pig adipose tissue-derived mesenchymal stem cell, gpBM-MSC guinea pig bone marrow-derived mesenchymal stem cell, MSC mesenchymal stem cell
Mentions: To examine the immunophenotype of guinea pig MSCs, cells were analysed by flow cytometry for the presence of positive MSC markers CD29, CD44, CD73 and CD90 and negative MSC markers CD34 (hematopoietic progenitors) and CD45 (leukocyte common antigen). Positive expression of CD29 was observed in both gpBM-MSCs (95.1 %) and gpAT-MSCs (97.6 %). There was differential expression of the CD73 antigen such that 85.0 % of gpBM-MSCs expressed the antigen compared with negligible expression in gpAT-MSCs (0.2 %) (Fig. 1a). Negligible expression of the remaining surface markers was observed in both cell lines (gpBM-MSC: CD44, 1.9 %; CD90, 1.5 %; CD34, 4.4 %; CD45, 5.5 %. gpAT-MSC: CD44, 0.2 %; CD90, 0.2 %; CD34, 0.2 %; CD45, 0.7 %; data not shown).Fig. 1

Bottom Line: This study aims to isolate and characterise guinea pig MSCs and then test their therapeutic potential for the treatment of enteric neuropathy associated with intestinal inflammation.MSCs from both sources secreted TGF-β1 which exerted neuroprotective effects in vitro.In vitro characteristics of MSCs cannot be extrapolated to their therapeutic efficacy.

View Article: PubMed Central - PubMed

Affiliation: Centre for Chronic Disease, College of Health and Biomedicine, Western Centre for Health, Research and Education, Sunshine Hospital, 176 Furlong road, Melbourne, 3021, Victoria, Australia. rhian.stavely@live.vu.edu.au.

ABSTRACT

Background: The use of mesenchymal stem cells (MSCs) to treat inflammatory bowel disease (IBD) is of great interest because of their immunomodulatory properties. Damage to the enteric nervous system (ENS) is implicated in IBD pathophysiology and disease progression. The most commonly used model to study inflammation-induced changes to the ENS is 2,4,6-trinitrobenzene-sulfonate acid (TNBS)-induced colitis in guinea pigs; however, no studies using guinea pig MSCs in colitis have been performed. This study aims to isolate and characterise guinea pig MSCs and then test their therapeutic potential for the treatment of enteric neuropathy associated with intestinal inflammation.

Methods: MSCs from guinea pig bone marrow and adipose tissue were isolated and characterised in vitro. In in vivo experiments, guinea pigs received either TNBS for the induction of colitis or sham treatment by enema. MSCs were administered at a dose of 1 × 10(6) cells via enema 3 h after the induction of colitis. Colon tissues were collected 24 and 72 h after TNBS administration to assess the level of inflammation and damage to the ENS. The secretion of transforming growth factor-β1 (TGF-β1) was analysed in MSC conditioned medium by flow cytometry.

Results: Cells isolated from both sources were adherent to plastic, multipotent and expressed some human MSC surface markers. In vitro characterisation revealed distinct differences in growth kinetics, clonogenicity and cell morphology between MSC types. In an in vivo model of TNBS-induced colitis, guinea pig bone marrow MSCs were comparatively more efficacious than adipose tissue MSCs in attenuating weight loss, colonic tissue damage and leukocyte infiltration into the mucosa and myenteric plexus. MSCs from both sources were equally neuroprotective in the amelioration of enteric neuronal loss and changes to the neurochemical coding of neuronal subpopulations. MSCs from both sources secreted TGF-β1 which exerted neuroprotective effects in vitro.

Conclusions: This study is the first evaluating the functional capacity of guinea pig bone marrow and adipose tissue-derived MSCs and providing evidence of their neuroprotective value in an animal model of colitis. In vitro characteristics of MSCs cannot be extrapolated to their therapeutic efficacy. TGF-β1 released by both types of MSCs might have contributed to the attenuation of enteric neuropathy associated with colitis.

No MeSH data available.


Related in: MedlinePlus