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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

MiR-140-5p Regulates CXCL12 in OM-MSCs(A) CXCL12 quantification in OM-MSC-CM, BM-MSC-CM, CD271-FT-CM, and fibroblast (FB)-CM. CXCL12 levels were significantly higher in OM-MSC-CM compared with all other cell types (n = 4 patient samples, mean ± SEM, ∗p < 0.05 determined by one-way ANOVA, Tukey's multiple comparison).(B and C) qPCR analysis of OM- and BM-MSCs transfected with miR-140-5p antagomir or mimic, a scrambled control, and dH2O only. MiR-140-5p expression was significantly greater in the mimic compared with controls (n = 3 patient samples, mean ± SEM, ∗∗∗p < 0.001, one-way ANOVA, Tukey's multiple comparison). BM-MSCs transfected with the mimic significantly increased levels from control (n = 3 patient samples, mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).(D) OM-MSC transfection with the miR-140-5p mimic caused significantly lower levels of CXCL12 mRNA compared with controls (n = 3 patient samples, mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).(E) Antagomir induced a significant increase in CXCL12 mRNA in BM-MSCs compared with control levels (n = 3 patient samples, mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).
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fig2: MiR-140-5p Regulates CXCL12 in OM-MSCs(A) CXCL12 quantification in OM-MSC-CM, BM-MSC-CM, CD271-FT-CM, and fibroblast (FB)-CM. CXCL12 levels were significantly higher in OM-MSC-CM compared with all other cell types (n = 4 patient samples, mean ± SEM, ∗p < 0.05 determined by one-way ANOVA, Tukey's multiple comparison).(B and C) qPCR analysis of OM- and BM-MSCs transfected with miR-140-5p antagomir or mimic, a scrambled control, and dH2O only. MiR-140-5p expression was significantly greater in the mimic compared with controls (n = 3 patient samples, mean ± SEM, ∗∗∗p < 0.001, one-way ANOVA, Tukey's multiple comparison). BM-MSCs transfected with the mimic significantly increased levels from control (n = 3 patient samples, mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).(D) OM-MSC transfection with the miR-140-5p mimic caused significantly lower levels of CXCL12 mRNA compared with controls (n = 3 patient samples, mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).(E) Antagomir induced a significant increase in CXCL12 mRNA in BM-MSCs compared with control levels (n = 3 patient samples, mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).

Mentions: GeneGo MetaCore analysis revealing high-confidence mRNA targets for miR-140-5p suggested SDF-1 (referred to as CXCL12 hereafter) secretion may be differentially regulated between the MSCs. Multiplex analysis of the conditioned media (CM) collected from OM- and BM-MSCs was performed with fibroblast (FB) and CD271-FT-CM used as comparisons (CM derived from n = 4 patient samples). Since we have previously shown that both FB-CM and CD271-FT-CM do not promote myelination (Lindsay et al., 2013), these were considered an appropriate comparison to ensure secreted factors were specifically generated from OM-MSCs (see Table 1); 13 were not detected and nine were secreted to equivalent levels across all groups (CCL1, CCL2, CCL3, CCL8, CX3CL1, G-CSF, CXCL10, SCF, and VEGF). CCL11 was significantly higher within both OM-MSC-CM and CD271-FT-CM compared with BM-MSC-CM and FB-CM (p < 0.05, all comparisons), suggesting the expression of a tissue-specific chemokine rather than MSC specific. CCL13 was significantly lower within FB-CM compared with all other cell type CM (p < 0.05, all comparisons), however, it was equivalently expressed within OM-MSCs, BM-MSCs, and CD271-FT-CM. CCL5 was markedly increased in CD271-FT-CM (p < 0.01). CXCL12 was the only cytokine present in significantly greater quantities in OM-MSC-CM compared with either BM-MSC-CM (p < 0.01), CD271-FT-CM (p < 0.01), or FB-CM (p < 0.05, Figure 2A). In addition, the neurotrophic factors BDNF, NT3, NT4/5, and NGF were assayed in BM- and OM-MSC-CM (n = 3 patient samples for both). NT3 was undetectable, and both cell types secreted equivalent low levels of BDNF (OM-MSC-CM, 19.2 ± 5.3 pg/ml; BM-MSC-CM, 11.9 ± 4.83 pg/ml) and NT4/5 (OM-MSC-CM, 21.9 ± 0.3 pg/ml; BM-MSC-CM, 20.0 ± 0.7 pg/ml). OM-MSC-CM contained significantly higher levels of NGF (33.8 ± 6.8 pg/ml) compared with BM-MSC-CM (1.2 ± 0.6 pg/ml).


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)

MiR-140-5p Regulates CXCL12 in OM-MSCs(A) CXCL12 quantification in OM-MSC-CM, BM-MSC-CM, CD271-FT-CM, and fibroblast (FB)-CM. CXCL12 levels were significantly higher in OM-MSC-CM compared with all other cell types (n = 4 patient samples, mean ± SEM, ∗p < 0.05 determined by one-way ANOVA, Tukey's multiple comparison).(B and C) qPCR analysis of OM- and BM-MSCs transfected with miR-140-5p antagomir or mimic, a scrambled control, and dH2O only. MiR-140-5p expression was significantly greater in the mimic compared with controls (n = 3 patient samples, mean ± SEM, ∗∗∗p < 0.001, one-way ANOVA, Tukey's multiple comparison). BM-MSCs transfected with the mimic significantly increased levels from control (n = 3 patient samples, mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).(D) OM-MSC transfection with the miR-140-5p mimic caused significantly lower levels of CXCL12 mRNA compared with controls (n = 3 patient samples, mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).(E) Antagomir induced a significant increase in CXCL12 mRNA in BM-MSCs compared with control levels (n = 3 patient samples, mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).
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fig2: MiR-140-5p Regulates CXCL12 in OM-MSCs(A) CXCL12 quantification in OM-MSC-CM, BM-MSC-CM, CD271-FT-CM, and fibroblast (FB)-CM. CXCL12 levels were significantly higher in OM-MSC-CM compared with all other cell types (n = 4 patient samples, mean ± SEM, ∗p < 0.05 determined by one-way ANOVA, Tukey's multiple comparison).(B and C) qPCR analysis of OM- and BM-MSCs transfected with miR-140-5p antagomir or mimic, a scrambled control, and dH2O only. MiR-140-5p expression was significantly greater in the mimic compared with controls (n = 3 patient samples, mean ± SEM, ∗∗∗p < 0.001, one-way ANOVA, Tukey's multiple comparison). BM-MSCs transfected with the mimic significantly increased levels from control (n = 3 patient samples, mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).(D) OM-MSC transfection with the miR-140-5p mimic caused significantly lower levels of CXCL12 mRNA compared with controls (n = 3 patient samples, mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).(E) Antagomir induced a significant increase in CXCL12 mRNA in BM-MSCs compared with control levels (n = 3 patient samples, mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).
Mentions: GeneGo MetaCore analysis revealing high-confidence mRNA targets for miR-140-5p suggested SDF-1 (referred to as CXCL12 hereafter) secretion may be differentially regulated between the MSCs. Multiplex analysis of the conditioned media (CM) collected from OM- and BM-MSCs was performed with fibroblast (FB) and CD271-FT-CM used as comparisons (CM derived from n = 4 patient samples). Since we have previously shown that both FB-CM and CD271-FT-CM do not promote myelination (Lindsay et al., 2013), these were considered an appropriate comparison to ensure secreted factors were specifically generated from OM-MSCs (see Table 1); 13 were not detected and nine were secreted to equivalent levels across all groups (CCL1, CCL2, CCL3, CCL8, CX3CL1, G-CSF, CXCL10, SCF, and VEGF). CCL11 was significantly higher within both OM-MSC-CM and CD271-FT-CM compared with BM-MSC-CM and FB-CM (p < 0.05, all comparisons), suggesting the expression of a tissue-specific chemokine rather than MSC specific. CCL13 was significantly lower within FB-CM compared with all other cell type CM (p < 0.05, all comparisons), however, it was equivalently expressed within OM-MSCs, BM-MSCs, and CD271-FT-CM. CCL5 was markedly increased in CD271-FT-CM (p < 0.01). CXCL12 was the only cytokine present in significantly greater quantities in OM-MSC-CM compared with either BM-MSC-CM (p < 0.01), CD271-FT-CM (p < 0.01), or FB-CM (p < 0.05, Figure 2A). In addition, the neurotrophic factors BDNF, NT3, NT4/5, and NGF were assayed in BM- and OM-MSC-CM (n = 3 patient samples for both). NT3 was undetectable, and both cell types secreted equivalent low levels of BDNF (OM-MSC-CM, 19.2 ± 5.3 pg/ml; BM-MSC-CM, 11.9 ± 4.83 pg/ml) and NT4/5 (OM-MSC-CM, 21.9 ± 0.3 pg/ml; BM-MSC-CM, 20.0 ± 0.7 pg/ml). OM-MSC-CM contained significantly higher levels of NGF (33.8 ± 6.8 pg/ml) compared with BM-MSC-CM (1.2 ± 0.6 pg/ml).

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