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Isolation of adipose and bone marrow mesenchymal stem cells using CD29 and CD90 modifies their capacity for osteogenic and adipogenic differentiation.

Davies OG, Cooper PR, Shelton RM, Smith AJ, Scheven BA - J Tissue Eng (2015)

Bottom Line: Maintaining cells in fluorescence-activated cell sorting buffer did not affect adipose-derived cell viability, but a significant (p < 0.05) reduction was found in bone marrow-derived cell viability.Additionally, CD29(+)/CD90(+) selection was associated with a significant decrease in the expression of Lin28, Sox2, Nanog and CD73 in adipose-derived cell cultures, whereas differences in stem cell-associated gene expression were not observed in sorted bone marrow-derived cell cultures.In summary, this study demonstrated that fluorescence-activated cell sorting had differential effects on adipose-derived cells and bone marrow-derived cells, and both CD29(+)/CD90(+) cells displayed a significantly reduced capacity for osteogenic/adipogenic differentiation.

View Article: PubMed Central - PubMed

Affiliation: School of Dentistry, University of Birmingham, Birmingham, UK ; Wolfson School of Mechanical and Manufacturing Engineering, Loughborough University, Loughborough, UK.

ABSTRACT
Mesenchymal stem cells isolated from rats are frequently used for tissue engineering research. However, considerable differences have been identified between rat mesenchymal stem cells and those derived from humans, and no defined panel of markers currently exists for the isolation of these cells. The aim of this study was to examine the effects of cell sorting for CD29(+)/CD90(+) cells from rat adipose and bone marrow tissues on their differentiation and expression of stem cell-associated genes. Flow cytometry showed 66% and 78% CD29(+)/CD90(+) positivity within passage 1 of adipose and bone marrow cultures, respectively. CD29(+)/CD90(+) cells showed a reduction in both osteogenic and adipogenic differentiation when compared with unsorted cells, as determined by alizarin red and Oil Red-O staining, respectively. These findings could not entirely be explained by fluorescence-activated cell sorting-induced cell injury as sort recovery was only modestly affected in adipose-derived cells. Maintaining cells in fluorescence-activated cell sorting buffer did not affect adipose-derived cell viability, but a significant (p < 0.05) reduction was found in bone marrow-derived cell viability. Additionally, CD29(+)/CD90(+) selection was associated with a significant decrease in the expression of Lin28, Sox2, Nanog and CD73 in adipose-derived cell cultures, whereas differences in stem cell-associated gene expression were not observed in sorted bone marrow-derived cell cultures. In summary, this study demonstrated that fluorescence-activated cell sorting had differential effects on adipose-derived cells and bone marrow-derived cells, and both CD29(+)/CD90(+) cells displayed a significantly reduced capacity for osteogenic/adipogenic differentiation. In conclusion, we identify that maintaining heterogeneity within the mesenchymal stem cell population may be important for optimal differentiation.

No MeSH data available.


Related in: MedlinePlus

Proportion of CD29+, CD90+ and CD29+/CD90+ cells within (a) passage 0 and (b) passage 1 ADC and BMDC cultures. Percentage positivity was analysed within the total cell population. Non-viable cells were excluded using propidium iodide staining. Data were analysed using Student’s t-test.*p < 0.05, n = 5.
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fig1-2041731415592356: Proportion of CD29+, CD90+ and CD29+/CD90+ cells within (a) passage 0 and (b) passage 1 ADC and BMDC cultures. Percentage positivity was analysed within the total cell population. Non-viable cells were excluded using propidium iodide staining. Data were analysed using Student’s t-test.*p < 0.05, n = 5.

Mentions: The total percentage of CD29+, CD90+ and CD29+/CD90+ cells within the whole population of adipose and bone marrow tissues was analysed using FACS (Figure 1). Freshly isolated (P0) ADC cultures contained significantly (p < 0.05) higher numbers of CD29+, CD90+ and CD29+/CD90+ cells than BMDC cultures (ADCs: 91% CD29+, 57% CD90+ and 51% CD29+/CD90+; BMDCs: 64% CD29+, 36% CD90+ and 28% CD29+/CD90+). No significant (p < 0.05) differences were identified between ADCs and BMDCs at P1 (ADCs: 98% CD29+, 73% CD90+ and 66% CD29+/CD90+; BMDCs: 99% CD29+, 84% CD90+ and 78% CD29+/CD90+).


Isolation of adipose and bone marrow mesenchymal stem cells using CD29 and CD90 modifies their capacity for osteogenic and adipogenic differentiation.

Davies OG, Cooper PR, Shelton RM, Smith AJ, Scheven BA - J Tissue Eng (2015)

Proportion of CD29+, CD90+ and CD29+/CD90+ cells within (a) passage 0 and (b) passage 1 ADC and BMDC cultures. Percentage positivity was analysed within the total cell population. Non-viable cells were excluded using propidium iodide staining. Data were analysed using Student’s t-test.*p < 0.05, n = 5.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2 - License 3
Show All Figures
getmorefigures.php?uid=PMC4555348&req=5

fig1-2041731415592356: Proportion of CD29+, CD90+ and CD29+/CD90+ cells within (a) passage 0 and (b) passage 1 ADC and BMDC cultures. Percentage positivity was analysed within the total cell population. Non-viable cells were excluded using propidium iodide staining. Data were analysed using Student’s t-test.*p < 0.05, n = 5.
Mentions: The total percentage of CD29+, CD90+ and CD29+/CD90+ cells within the whole population of adipose and bone marrow tissues was analysed using FACS (Figure 1). Freshly isolated (P0) ADC cultures contained significantly (p < 0.05) higher numbers of CD29+, CD90+ and CD29+/CD90+ cells than BMDC cultures (ADCs: 91% CD29+, 57% CD90+ and 51% CD29+/CD90+; BMDCs: 64% CD29+, 36% CD90+ and 28% CD29+/CD90+). No significant (p < 0.05) differences were identified between ADCs and BMDCs at P1 (ADCs: 98% CD29+, 73% CD90+ and 66% CD29+/CD90+; BMDCs: 99% CD29+, 84% CD90+ and 78% CD29+/CD90+).

Bottom Line: Maintaining cells in fluorescence-activated cell sorting buffer did not affect adipose-derived cell viability, but a significant (p < 0.05) reduction was found in bone marrow-derived cell viability.Additionally, CD29(+)/CD90(+) selection was associated with a significant decrease in the expression of Lin28, Sox2, Nanog and CD73 in adipose-derived cell cultures, whereas differences in stem cell-associated gene expression were not observed in sorted bone marrow-derived cell cultures.In summary, this study demonstrated that fluorescence-activated cell sorting had differential effects on adipose-derived cells and bone marrow-derived cells, and both CD29(+)/CD90(+) cells displayed a significantly reduced capacity for osteogenic/adipogenic differentiation.

View Article: PubMed Central - PubMed

Affiliation: School of Dentistry, University of Birmingham, Birmingham, UK ; Wolfson School of Mechanical and Manufacturing Engineering, Loughborough University, Loughborough, UK.

ABSTRACT
Mesenchymal stem cells isolated from rats are frequently used for tissue engineering research. However, considerable differences have been identified between rat mesenchymal stem cells and those derived from humans, and no defined panel of markers currently exists for the isolation of these cells. The aim of this study was to examine the effects of cell sorting for CD29(+)/CD90(+) cells from rat adipose and bone marrow tissues on their differentiation and expression of stem cell-associated genes. Flow cytometry showed 66% and 78% CD29(+)/CD90(+) positivity within passage 1 of adipose and bone marrow cultures, respectively. CD29(+)/CD90(+) cells showed a reduction in both osteogenic and adipogenic differentiation when compared with unsorted cells, as determined by alizarin red and Oil Red-O staining, respectively. These findings could not entirely be explained by fluorescence-activated cell sorting-induced cell injury as sort recovery was only modestly affected in adipose-derived cells. Maintaining cells in fluorescence-activated cell sorting buffer did not affect adipose-derived cell viability, but a significant (p < 0.05) reduction was found in bone marrow-derived cell viability. Additionally, CD29(+)/CD90(+) selection was associated with a significant decrease in the expression of Lin28, Sox2, Nanog and CD73 in adipose-derived cell cultures, whereas differences in stem cell-associated gene expression were not observed in sorted bone marrow-derived cell cultures. In summary, this study demonstrated that fluorescence-activated cell sorting had differential effects on adipose-derived cells and bone marrow-derived cells, and both CD29(+)/CD90(+) cells displayed a significantly reduced capacity for osteogenic/adipogenic differentiation. In conclusion, we identify that maintaining heterogeneity within the mesenchymal stem cell population may be important for optimal differentiation.

No MeSH data available.


Related in: MedlinePlus