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Comparative miRNA-Based Fingerprinting Reveals Biological Differences in Human Olfactory Mucosa- and Bone-Marrow-Derived Mesenchymal Stromal Cells.

Lindsay SL, Johnstone SA, McGrath MA, Mallinson D, Barnett SC - Stem Cell Reports (2016)

Bottom Line: Previously we reported that nestin-positive human mesenchymal stromal cells (MSCs) derived from the olfactory mucosa (OM) enhanced CNS myelination in vitro to a greater extent than bone-marrow-derived MSCs (BM-MSCs). miRNA-based fingerprinting revealed the two MSCs were 64% homologous, with 26 miRNAs differentially expressed.The lower expression of miR-140-5p in OM-MSCs correlated with higher secretion of CXCL12 compared with BM-MSCs.Nestin-positive OM-MSCs could therefore offer a cell transplantation alternative for CNS repair, should these biological behaviors be translated in vivo.

View Article: PubMed Central - PubMed

Affiliation: Institute of Infection, Inflammation and Immunity, Glasgow Biomedical Research Centre, University of Glasgow, Sir Graeme Davies Building, 120 University Place, Glasgow G12 8TA, UK.

No MeSH data available.


Related in: MedlinePlus

CXCL12 Promotes In Vitro CNS Myelination which is Regulated by miR-140-5p(A) Co-cultures stained for myelin (green PLP) and axons (red SMI-31) treated with OM-MSC-CM, OM-MSC-CM in the presence of antibody to CXCL12 (OM-MSC-CM + anti-CXCL12), or CXCR4 blocker (OM-MSC-CM + AMD3100), CXCL12 (100 ng/ml), or CXCL12 in the presence of AMD3100. Scale bar represents 100 μm.(B) CXCL12 and OM-MSC-CM increased myelination significantly compared with control levels (demarcated by the dotted line). AMD3100 blocker and anti-CXCL12 abolished the pro-myelinating effect of OM-MSC-CM and CXCL12 but did not affect myelination on their own (n = 4 patient samples, mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).(C) Representative images of co-cultures stained for myelin (green PLP) and axons (red SMI-31) treated with CM from BM- or OM-MSCs transfected with miRNA control, antagomir (140-5p Antag), or miR-140-5p mimic and miR-140-5p antagomir-derived CM in the presence of anti-CXCL12 (140-5p + Ab) or the blocker (140-5p + AMD3100). Scale bar represents 100 μm.(D) CM from BM-MSCs treated with miR-140-5p antagomir led to a significant increase in myelination compared with control (n = 6 patient samples for all). The antagomir-induced increase could be reduced to control levels in the presence of anti-CXCL12 or AMD3100 blocker (n = 3 patient samples for both). CM from BM-MSCs induced by the miR-140-5p mimic also did not promote myelination.(E) OM-MSC-CM collected after miR-140-5p mimic transfection significantly reduced the pro-myelinating effect of OM-MSCs while transfection with antagomir had no effect (n = 3 patient samples). In (D) and (E), mean ± SEM, ∗p < 0.05, ∗∗p < 0.01, one-way ANOVA, Tukey's multiple comparison.Dotted horizontal lines demarcate control levels, vertical dotted lines detail experimental comparisons made to control.
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fig3: CXCL12 Promotes In Vitro CNS Myelination which is Regulated by miR-140-5p(A) Co-cultures stained for myelin (green PLP) and axons (red SMI-31) treated with OM-MSC-CM, OM-MSC-CM in the presence of antibody to CXCL12 (OM-MSC-CM + anti-CXCL12), or CXCR4 blocker (OM-MSC-CM + AMD3100), CXCL12 (100 ng/ml), or CXCL12 in the presence of AMD3100. Scale bar represents 100 μm.(B) CXCL12 and OM-MSC-CM increased myelination significantly compared with control levels (demarcated by the dotted line). AMD3100 blocker and anti-CXCL12 abolished the pro-myelinating effect of OM-MSC-CM and CXCL12 but did not affect myelination on their own (n = 4 patient samples, mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).(C) Representative images of co-cultures stained for myelin (green PLP) and axons (red SMI-31) treated with CM from BM- or OM-MSCs transfected with miRNA control, antagomir (140-5p Antag), or miR-140-5p mimic and miR-140-5p antagomir-derived CM in the presence of anti-CXCL12 (140-5p + Ab) or the blocker (140-5p + AMD3100). Scale bar represents 100 μm.(D) CM from BM-MSCs treated with miR-140-5p antagomir led to a significant increase in myelination compared with control (n = 6 patient samples for all). The antagomir-induced increase could be reduced to control levels in the presence of anti-CXCL12 or AMD3100 blocker (n = 3 patient samples for both). CM from BM-MSCs induced by the miR-140-5p mimic also did not promote myelination.(E) OM-MSC-CM collected after miR-140-5p mimic transfection significantly reduced the pro-myelinating effect of OM-MSCs while transfection with antagomir had no effect (n = 3 patient samples). In (D) and (E), mean ± SEM, ∗p < 0.05, ∗∗p < 0.01, one-way ANOVA, Tukey's multiple comparison.Dotted horizontal lines demarcate control levels, vertical dotted lines detail experimental comparisons made to control.

Mentions: Our previous data suggest that OM-MSCs promoted in vitro CNS myelination via a secreted factor. Here we provide evidence that CXCL12 could be this factor. Myelinating CNS co-cultures were treated with CXCL12 (100 ng/ml), CXCL12 receptor CXCR4 blocker (AMD3100), OM-MSC-CM, as well as OM-MSC-CM treated with a neutralizing antibody to CXCL12 or AMD3100 (n = 4, all treatments and four different patient samples; Figures 3A and 3B). Media containing the CXCL12 neutralizing antibody or AMD3100 were used as controls. Exogenous CXCL12 significantly increased myelination almost 2-fold compared with controls (p < 0.05), which was blocked by AMD3100. The pro-myelinating effect of OM-MSC-CM (p < 0.05) was also reduced by AMD3100 and when treated with the neutralizing antibody to CXCL12. These data indicate that CXCL12 is at least partially responsible for the pro-myelinating effect of OM-MSCs.


Comparative miRNA-Based Fingerprinting Reveals Biological Differences in Human Olfactory Mucosa- and Bone-Marrow-Derived Mesenchymal Stromal Cells.

Lindsay SL, Johnstone SA, McGrath MA, Mallinson D, Barnett SC - Stem Cell Reports (2016)

CXCL12 Promotes In Vitro CNS Myelination which is Regulated by miR-140-5p(A) Co-cultures stained for myelin (green PLP) and axons (red SMI-31) treated with OM-MSC-CM, OM-MSC-CM in the presence of antibody to CXCL12 (OM-MSC-CM + anti-CXCL12), or CXCR4 blocker (OM-MSC-CM + AMD3100), CXCL12 (100 ng/ml), or CXCL12 in the presence of AMD3100. Scale bar represents 100 μm.(B) CXCL12 and OM-MSC-CM increased myelination significantly compared with control levels (demarcated by the dotted line). AMD3100 blocker and anti-CXCL12 abolished the pro-myelinating effect of OM-MSC-CM and CXCL12 but did not affect myelination on their own (n = 4 patient samples, mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).(C) Representative images of co-cultures stained for myelin (green PLP) and axons (red SMI-31) treated with CM from BM- or OM-MSCs transfected with miRNA control, antagomir (140-5p Antag), or miR-140-5p mimic and miR-140-5p antagomir-derived CM in the presence of anti-CXCL12 (140-5p + Ab) or the blocker (140-5p + AMD3100). Scale bar represents 100 μm.(D) CM from BM-MSCs treated with miR-140-5p antagomir led to a significant increase in myelination compared with control (n = 6 patient samples for all). The antagomir-induced increase could be reduced to control levels in the presence of anti-CXCL12 or AMD3100 blocker (n = 3 patient samples for both). CM from BM-MSCs induced by the miR-140-5p mimic also did not promote myelination.(E) OM-MSC-CM collected after miR-140-5p mimic transfection significantly reduced the pro-myelinating effect of OM-MSCs while transfection with antagomir had no effect (n = 3 patient samples). In (D) and (E), mean ± SEM, ∗p < 0.05, ∗∗p < 0.01, one-way ANOVA, Tukey's multiple comparison.Dotted horizontal lines demarcate control levels, vertical dotted lines detail experimental comparisons made to control.
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fig3: CXCL12 Promotes In Vitro CNS Myelination which is Regulated by miR-140-5p(A) Co-cultures stained for myelin (green PLP) and axons (red SMI-31) treated with OM-MSC-CM, OM-MSC-CM in the presence of antibody to CXCL12 (OM-MSC-CM + anti-CXCL12), or CXCR4 blocker (OM-MSC-CM + AMD3100), CXCL12 (100 ng/ml), or CXCL12 in the presence of AMD3100. Scale bar represents 100 μm.(B) CXCL12 and OM-MSC-CM increased myelination significantly compared with control levels (demarcated by the dotted line). AMD3100 blocker and anti-CXCL12 abolished the pro-myelinating effect of OM-MSC-CM and CXCL12 but did not affect myelination on their own (n = 4 patient samples, mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).(C) Representative images of co-cultures stained for myelin (green PLP) and axons (red SMI-31) treated with CM from BM- or OM-MSCs transfected with miRNA control, antagomir (140-5p Antag), or miR-140-5p mimic and miR-140-5p antagomir-derived CM in the presence of anti-CXCL12 (140-5p + Ab) or the blocker (140-5p + AMD3100). Scale bar represents 100 μm.(D) CM from BM-MSCs treated with miR-140-5p antagomir led to a significant increase in myelination compared with control (n = 6 patient samples for all). The antagomir-induced increase could be reduced to control levels in the presence of anti-CXCL12 or AMD3100 blocker (n = 3 patient samples for both). CM from BM-MSCs induced by the miR-140-5p mimic also did not promote myelination.(E) OM-MSC-CM collected after miR-140-5p mimic transfection significantly reduced the pro-myelinating effect of OM-MSCs while transfection with antagomir had no effect (n = 3 patient samples). In (D) and (E), mean ± SEM, ∗p < 0.05, ∗∗p < 0.01, one-way ANOVA, Tukey's multiple comparison.Dotted horizontal lines demarcate control levels, vertical dotted lines detail experimental comparisons made to control.
Mentions: Our previous data suggest that OM-MSCs promoted in vitro CNS myelination via a secreted factor. Here we provide evidence that CXCL12 could be this factor. Myelinating CNS co-cultures were treated with CXCL12 (100 ng/ml), CXCL12 receptor CXCR4 blocker (AMD3100), OM-MSC-CM, as well as OM-MSC-CM treated with a neutralizing antibody to CXCL12 or AMD3100 (n = 4, all treatments and four different patient samples; Figures 3A and 3B). Media containing the CXCL12 neutralizing antibody or AMD3100 were used as controls. Exogenous CXCL12 significantly increased myelination almost 2-fold compared with controls (p < 0.05), which was blocked by AMD3100. The pro-myelinating effect of OM-MSC-CM (p < 0.05) was also reduced by AMD3100 and when treated with the neutralizing antibody to CXCL12. These data indicate that CXCL12 is at least partially responsible for the pro-myelinating effect of OM-MSCs.

Bottom Line: Previously we reported that nestin-positive human mesenchymal stromal cells (MSCs) derived from the olfactory mucosa (OM) enhanced CNS myelination in vitro to a greater extent than bone-marrow-derived MSCs (BM-MSCs). miRNA-based fingerprinting revealed the two MSCs were 64% homologous, with 26 miRNAs differentially expressed.The lower expression of miR-140-5p in OM-MSCs correlated with higher secretion of CXCL12 compared with BM-MSCs.Nestin-positive OM-MSCs could therefore offer a cell transplantation alternative for CNS repair, should these biological behaviors be translated in vivo.

View Article: PubMed Central - PubMed

Affiliation: Institute of Infection, Inflammation and Immunity, Glasgow Biomedical Research Centre, University of Glasgow, Sir Graeme Davies Building, 120 University Place, Glasgow G12 8TA, UK.

No MeSH data available.


Related in: MedlinePlus