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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

Differentiation potential of guinea pig MSCs. GpBM-MSCs and gpAT-MSCs cultured without (a, b) and with (a′, b′) adipogenesis differentiation medium for 14 days and stained with Oil red O. Scale bar = 50 μm. GpBM-MSCs and gpAT-MSCs cultured without (c, d) and with (c′, d′) osteogenesis differentiation medium for 21 days and stained with Alizarin red S. Scale bar = 200 μm. Alcian blue stained cross-sections of chondrogenic pellets formed by gpBM-MSCs (e) and gpAT-MSCs (f) after 14 days in chondrogenic differentiation medium. Scale bar = 50 μm. 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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Fig2: Differentiation potential of guinea pig MSCs. GpBM-MSCs and gpAT-MSCs cultured without (a, b) and with (a′, b′) adipogenesis differentiation medium for 14 days and stained with Oil red O. Scale bar = 50 μm. GpBM-MSCs and gpAT-MSCs cultured without (c, d) and with (c′, d′) osteogenesis differentiation medium for 21 days and stained with Alizarin red S. Scale bar = 200 μm. Alcian blue stained cross-sections of chondrogenic pellets formed by gpBM-MSCs (e) and gpAT-MSCs (f) after 14 days in chondrogenic differentiation medium. Scale bar = 50 μm. 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: Both bone marrow- and adipose tissue-derived MSCs grew in monolayer culture, adhered to plastic, proliferated and were typical of MSC appearance (Fig. 1b, b′). The clonogenicity of MSCs was compared via a CFU-f assay. The yield of colony-forming units (>50 cells per colony [73]) of gpAT-MSCs (17.0 ± 1.3 %) was higher than that of gpBM-MSCs (1.3 ± 0.5 %, P <0.0001) after 2 weeks in culture (Fig. 1c, c′, d, n = 4 independent cultures per group). To assess the multipotent potential of MSCs, cells were cultured in specialised media to induce adipogenic, osteogenic and chondrogenic differentiation. Lipid vacuoles were present in MSCs cultured in adipogenesis differentiation medium when stained with Oil red O indicative of successful differentiation into adipocytes (Fig. 2a–b′). MSCs cultured in osteogenesis differentiation medium were stained with Alizarin red S, confirming successful differentiation into osteocytes (Fig. 2c–d′). MSCs seeded as micromass cultures only formed pellets in chondrogenesis differentiation medium. Chondrogenic differentiation was confirmed in the pellets by Alcian blue staining of cross-sections (Fig. 2e, f).Fig. 2


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)

Differentiation potential of guinea pig MSCs. GpBM-MSCs and gpAT-MSCs cultured without (a, b) and with (a′, b′) adipogenesis differentiation medium for 14 days and stained with Oil red O. Scale bar = 50 μm. GpBM-MSCs and gpAT-MSCs cultured without (c, d) and with (c′, d′) osteogenesis differentiation medium for 21 days and stained with Alizarin red S. Scale bar = 200 μm. Alcian blue stained cross-sections of chondrogenic pellets formed by gpBM-MSCs (e) and gpAT-MSCs (f) after 14 days in chondrogenic differentiation medium. Scale bar = 50 μm. 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

Fig2: Differentiation potential of guinea pig MSCs. GpBM-MSCs and gpAT-MSCs cultured without (a, b) and with (a′, b′) adipogenesis differentiation medium for 14 days and stained with Oil red O. Scale bar = 50 μm. GpBM-MSCs and gpAT-MSCs cultured without (c, d) and with (c′, d′) osteogenesis differentiation medium for 21 days and stained with Alizarin red S. Scale bar = 200 μm. Alcian blue stained cross-sections of chondrogenic pellets formed by gpBM-MSCs (e) and gpAT-MSCs (f) after 14 days in chondrogenic differentiation medium. Scale bar = 50 μm. 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: Both bone marrow- and adipose tissue-derived MSCs grew in monolayer culture, adhered to plastic, proliferated and were typical of MSC appearance (Fig. 1b, b′). The clonogenicity of MSCs was compared via a CFU-f assay. The yield of colony-forming units (>50 cells per colony [73]) of gpAT-MSCs (17.0 ± 1.3 %) was higher than that of gpBM-MSCs (1.3 ± 0.5 %, P <0.0001) after 2 weeks in culture (Fig. 1c, c′, d, n = 4 independent cultures per group). To assess the multipotent potential of MSCs, cells were cultured in specialised media to induce adipogenic, osteogenic and chondrogenic differentiation. Lipid vacuoles were present in MSCs cultured in adipogenesis differentiation medium when stained with Oil red O indicative of successful differentiation into adipocytes (Fig. 2a–b′). MSCs cultured in osteogenesis differentiation medium were stained with Alizarin red S, confirming successful differentiation into osteocytes (Fig. 2c–d′). MSCs seeded as micromass cultures only formed pellets in chondrogenesis differentiation medium. Chondrogenic differentiation was confirmed in the pellets by Alcian blue staining of cross-sections (Fig. 2e, f).Fig. 2

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