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In Vitro Differentiation of First Trimester Human Umbilical Cord Perivascular Cells into Contracting Cardiomyocyte-Like Cells.

Szaraz P, Librach M, Maghen L, Iqbal F, Barretto TA, Kenigsberg S, Gauthier-Fisher A, Librach CL - Stem Cells Int (2016)

Bottom Line: However the optimal cell type providing significant cardiac regeneration after MI is yet to be found.Based on the expression of cardiomyocyte markers (cTnT, MYH6, SIRPA, and CX43) FTM and term HUCPVCs achieved significantly increased cardiomyogenic differentiation compared to bone marrow MSCs, while their immunogenicity remained significantly lower as indicated by HLA-A and HLA-G expression and susceptibility to T cell mediated cytotoxicity.Our results indicate that young FTM HUCPVCs have superior cardiomyogenic potential coupled with beneficial immunogenic properties when compared to MSCs of older tissue sources, suggesting that in vitro predifferentiation could be a potential strategy to increase their effectiveness in vivo.

View Article: PubMed Central - PubMed

Affiliation: Create Fertility Centre, Toronto, ON, Canada M5G 1N8; Institute of Medical Sciences, University of Toronto, Toronto, ON, Canada M5S 1A8.

ABSTRACT
Myocardial infarction (MI) causes an extensive loss of heart muscle cells and leads to congestive heart disease (CAD), the leading cause of mortality and morbidity worldwide. Mesenchymal stromal cell- (MSC-) based cell therapy is a promising option to replace invasive interventions. However the optimal cell type providing significant cardiac regeneration after MI is yet to be found. The aim of our study was to investigate the cardiomyogenic differentiation potential of first trimester human umbilical cord perivascular cells (FTM HUCPVCs), a novel, young source of immunoprivileged mesenchymal stromal cells. Based on the expression of cardiomyocyte markers (cTnT, MYH6, SIRPA, and CX43) FTM and term HUCPVCs achieved significantly increased cardiomyogenic differentiation compared to bone marrow MSCs, while their immunogenicity remained significantly lower as indicated by HLA-A and HLA-G expression and susceptibility to T cell mediated cytotoxicity. When applying aggregate-based differentiation, FTM HUCPVCs showed increased aggregate formation potential and generated contracting cells within 1 week of coculture, making them the first MSC type with this ability. Our results indicate that young FTM HUCPVCs have superior cardiomyogenic potential coupled with beneficial immunogenic properties when compared to MSCs of older tissue sources, suggesting that in vitro predifferentiation could be a potential strategy to increase their effectiveness in vivo.

No MeSH data available.


Related in: MedlinePlus

Immunological characteristics of hMSCs before and after cardiac differentiation in vitro. (a–c) FC analysis of HLA-A and HLA-G expression in (a) undifferentiated, (b) direct coculture, and (c) aggregate coculture differentiated FTM and term HUCPVCs and bone marrow MSCs (BMSCs). Values expressed as % of overall human cell number in corresponding cultures. (d) Colorimetric analysis of human leukocyte induced LDH release after 72 h of incubation with FTM and term HUCPVCs and BMSCs sorted from cardiomyocyte cultures (direct cocultures) compared to undifferentiated MSC cultures. Values expressed as optical density (OD) of LDH converted substrate detected at 490 nm. Inner graph shows undifferentiated MSC values (y-axis 3x of main graph). Asterisk indicates statistically significant difference (p < 0.01, nFTM = 9, nterm = 9, nBMSC = 6).
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fig3: Immunological characteristics of hMSCs before and after cardiac differentiation in vitro. (a–c) FC analysis of HLA-A and HLA-G expression in (a) undifferentiated, (b) direct coculture, and (c) aggregate coculture differentiated FTM and term HUCPVCs and bone marrow MSCs (BMSCs). Values expressed as % of overall human cell number in corresponding cultures. (d) Colorimetric analysis of human leukocyte induced LDH release after 72 h of incubation with FTM and term HUCPVCs and BMSCs sorted from cardiomyocyte cultures (direct cocultures) compared to undifferentiated MSC cultures. Values expressed as optical density (OD) of LDH converted substrate detected at 490 nm. Inner graph shows undifferentiated MSC values (y-axis 3x of main graph). Asterisk indicates statistically significant difference (p < 0.01, nFTM = 9, nterm = 9, nBMSC = 6).

Mentions: In undifferentiated cultures, HLA-A levels were significantly lower for FTM (23.5% ± 15.2%) and term HUCPVCs (32.3% ± 23%) when compared to BMSCs (70.3% ± 8.1%, p < 0.01) (Figure 3(a)). The proportion of immune privilege-associated HLA-G positivity was significantly higher in FTM (18.2% ± 6.4%) and term HUCPVCs (14.7% ± 6%) cells when compared to BMSCs (4.2% ± 2.8) (p < 0.01) (Figure 3(b)). Cardiomyogenic differentiation in cocultures led to upregulation of HLA-A in all MSC types, but HLA-A expression remained significantly lower for FTM (40.8% ± 18.2%) and term HUCPVCs (57.8% ± 26.1%) in comparison to BMSCs (90.7% ± 7.8%) (p < 0.01) (Figure 3(a)). In aggregate cocultures, proportions of HLA-A-positive cells increased in term HUCPVCs (85.4% ± 3%) but remained significantly lower in FTM HUCPVCs (53.6% ± 22.8%) (p < 0.01). HLA-G levels remained significantly higher in both direct cocultures (FTM: 23.8% ± 6.2%, term: 21% ± 4.7%) and aggregate cocultures (FTM 42.8 ± 30.4%, term 29.6% ± 6%) when compared to BMSCs (3.2% ± 1.7% and 3.7% ± 0.7%, resp.) (p < 0.01) (Figure 3(c)).


In Vitro Differentiation of First Trimester Human Umbilical Cord Perivascular Cells into Contracting Cardiomyocyte-Like Cells.

Szaraz P, Librach M, Maghen L, Iqbal F, Barretto TA, Kenigsberg S, Gauthier-Fisher A, Librach CL - Stem Cells Int (2016)

Immunological characteristics of hMSCs before and after cardiac differentiation in vitro. (a–c) FC analysis of HLA-A and HLA-G expression in (a) undifferentiated, (b) direct coculture, and (c) aggregate coculture differentiated FTM and term HUCPVCs and bone marrow MSCs (BMSCs). Values expressed as % of overall human cell number in corresponding cultures. (d) Colorimetric analysis of human leukocyte induced LDH release after 72 h of incubation with FTM and term HUCPVCs and BMSCs sorted from cardiomyocyte cultures (direct cocultures) compared to undifferentiated MSC cultures. Values expressed as optical density (OD) of LDH converted substrate detected at 490 nm. Inner graph shows undifferentiated MSC values (y-axis 3x of main graph). Asterisk indicates statistically significant difference (p < 0.01, nFTM = 9, nterm = 9, nBMSC = 6).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig3: Immunological characteristics of hMSCs before and after cardiac differentiation in vitro. (a–c) FC analysis of HLA-A and HLA-G expression in (a) undifferentiated, (b) direct coculture, and (c) aggregate coculture differentiated FTM and term HUCPVCs and bone marrow MSCs (BMSCs). Values expressed as % of overall human cell number in corresponding cultures. (d) Colorimetric analysis of human leukocyte induced LDH release after 72 h of incubation with FTM and term HUCPVCs and BMSCs sorted from cardiomyocyte cultures (direct cocultures) compared to undifferentiated MSC cultures. Values expressed as optical density (OD) of LDH converted substrate detected at 490 nm. Inner graph shows undifferentiated MSC values (y-axis 3x of main graph). Asterisk indicates statistically significant difference (p < 0.01, nFTM = 9, nterm = 9, nBMSC = 6).
Mentions: In undifferentiated cultures, HLA-A levels were significantly lower for FTM (23.5% ± 15.2%) and term HUCPVCs (32.3% ± 23%) when compared to BMSCs (70.3% ± 8.1%, p < 0.01) (Figure 3(a)). The proportion of immune privilege-associated HLA-G positivity was significantly higher in FTM (18.2% ± 6.4%) and term HUCPVCs (14.7% ± 6%) cells when compared to BMSCs (4.2% ± 2.8) (p < 0.01) (Figure 3(b)). Cardiomyogenic differentiation in cocultures led to upregulation of HLA-A in all MSC types, but HLA-A expression remained significantly lower for FTM (40.8% ± 18.2%) and term HUCPVCs (57.8% ± 26.1%) in comparison to BMSCs (90.7% ± 7.8%) (p < 0.01) (Figure 3(a)). In aggregate cocultures, proportions of HLA-A-positive cells increased in term HUCPVCs (85.4% ± 3%) but remained significantly lower in FTM HUCPVCs (53.6% ± 22.8%) (p < 0.01). HLA-G levels remained significantly higher in both direct cocultures (FTM: 23.8% ± 6.2%, term: 21% ± 4.7%) and aggregate cocultures (FTM 42.8 ± 30.4%, term 29.6% ± 6%) when compared to BMSCs (3.2% ± 1.7% and 3.7% ± 0.7%, resp.) (p < 0.01) (Figure 3(c)).

Bottom Line: However the optimal cell type providing significant cardiac regeneration after MI is yet to be found.Based on the expression of cardiomyocyte markers (cTnT, MYH6, SIRPA, and CX43) FTM and term HUCPVCs achieved significantly increased cardiomyogenic differentiation compared to bone marrow MSCs, while their immunogenicity remained significantly lower as indicated by HLA-A and HLA-G expression and susceptibility to T cell mediated cytotoxicity.Our results indicate that young FTM HUCPVCs have superior cardiomyogenic potential coupled with beneficial immunogenic properties when compared to MSCs of older tissue sources, suggesting that in vitro predifferentiation could be a potential strategy to increase their effectiveness in vivo.

View Article: PubMed Central - PubMed

Affiliation: Create Fertility Centre, Toronto, ON, Canada M5G 1N8; Institute of Medical Sciences, University of Toronto, Toronto, ON, Canada M5S 1A8.

ABSTRACT
Myocardial infarction (MI) causes an extensive loss of heart muscle cells and leads to congestive heart disease (CAD), the leading cause of mortality and morbidity worldwide. Mesenchymal stromal cell- (MSC-) based cell therapy is a promising option to replace invasive interventions. However the optimal cell type providing significant cardiac regeneration after MI is yet to be found. The aim of our study was to investigate the cardiomyogenic differentiation potential of first trimester human umbilical cord perivascular cells (FTM HUCPVCs), a novel, young source of immunoprivileged mesenchymal stromal cells. Based on the expression of cardiomyocyte markers (cTnT, MYH6, SIRPA, and CX43) FTM and term HUCPVCs achieved significantly increased cardiomyogenic differentiation compared to bone marrow MSCs, while their immunogenicity remained significantly lower as indicated by HLA-A and HLA-G expression and susceptibility to T cell mediated cytotoxicity. When applying aggregate-based differentiation, FTM HUCPVCs showed increased aggregate formation potential and generated contracting cells within 1 week of coculture, making them the first MSC type with this ability. Our results indicate that young FTM HUCPVCs have superior cardiomyogenic potential coupled with beneficial immunogenic properties when compared to MSCs of older tissue sources, suggesting that in vitro predifferentiation could be a potential strategy to increase their effectiveness in vivo.

No MeSH data available.


Related in: MedlinePlus