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Improved adipogenic in vitro differentiation: comparison of different adipogenic cell culture media on human fat and bone stroma cells for fat tissue engineering.

Ghoniem AA, Açil Y, Wiltfang J, Gierloff M - Anat Cell Biol (2015)

Bottom Line: Our results revealed that only SAM was significantly (P<0.05) superior in the differentiation process in contrast to NAM for 10 days and 21 days.As well was SAM superior to differentiate the used cell lineages.The results suggest that the SAM should be established as a new standard medium for a more promising in vitro adipogenic differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral and Maxillofacial Surgery, Christian-Albrechts-University, Kiel, Germany.

ABSTRACT
To date there is no sufficient in vitro fat tissue engineering and a protocol has not been well established for this purpose. Therefore, we evaluated the in vitro influence of two different adipogenic growth media for their stimulation potential on different cell lineages to clearly define the most potent adipogenic growth media for future in vitro tissue engineering approaches. The samples for differentiation were composed of human adipogenic-derived stroma cells (hADSCs) and human bone marrow mesenchymal stroma cells (hMSCs). A normal adipogenic medium (NAM) and a specific adipogenic medium (SAM) were tested for their adipogenic stimulation potential. After 10 days and 21 days the relative gene expression was measured for the adipogenic marker genes PPARγ2, C/EBPα, FABP4, LPL, and GLUT4 detected through real time reverse transcriptase polymease chain reaction (RT-PCR). Other study variables were the comparison between NAM and SAM and between the used cells hADSCs and hMSCs. Additionally an Oil-Red staining was performed after 21 days. Our results revealed that only SAM was significantly (P<0.05) superior in the differentiation process in contrast to NAM for 10 days and 21 days. As well was SAM superior to differentiate the used cell lineages. This was evaluated by the detected marker genes PPARγ2, C/EBPα, FABP4, LPL, and GLUT4 through real time RT-PCR and by Oil-Red staining. In addition, the hMSCs proofed to be equal donor cells for adipogenic differentiation especially when stimulated by SAM. The results suggest that the SAM should be established as a new standard medium for a more promising in vitro adipogenic differentiation.

No MeSH data available.


Flowchart shows the allocation of adipogenic growth media (SAM and NAM) with each cell type (hADSC and hMSC). Arrows show up- and downregulation of the relative gene expression level. To get a further insight of relative expression level ratios please refer to Fig. 1. Significant differences where only detectable for SAM towards NAM. For further statistic exploration please refer to Table 4. hADSC, human adipogenic-derived stroma cell; hMSC, human bone marrow mesenchymal stroma cell; NAM, normal adipogenic medium; SAM, specific adipogenic medium.
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Figure 3: Flowchart shows the allocation of adipogenic growth media (SAM and NAM) with each cell type (hADSC and hMSC). Arrows show up- and downregulation of the relative gene expression level. To get a further insight of relative expression level ratios please refer to Fig. 1. Significant differences where only detectable for SAM towards NAM. For further statistic exploration please refer to Table 4. hADSC, human adipogenic-derived stroma cell; hMSC, human bone marrow mesenchymal stroma cell; NAM, normal adipogenic medium; SAM, specific adipogenic medium.

Mentions: At the second primer point at day 21 (t2), there were only measurable significant differences for LPL (P≤0.01) and FABP4 (P≤0.05). LPL's relative expression decreased and FABP4 stayed on a constant level. The marker genes PPARγ2 and GLUT4 showed an increased relative expression compared to primer point one (t1). In contrast C/EBPα showed a constant relative expression. For the PPARγ2, C/EBPα, and GLUT4 marker gene, there were no significant differences detectable between SAM and NAM (P>0.05) (Table 4, Figs. 2, 3).


Improved adipogenic in vitro differentiation: comparison of different adipogenic cell culture media on human fat and bone stroma cells for fat tissue engineering.

Ghoniem AA, Açil Y, Wiltfang J, Gierloff M - Anat Cell Biol (2015)

Flowchart shows the allocation of adipogenic growth media (SAM and NAM) with each cell type (hADSC and hMSC). Arrows show up- and downregulation of the relative gene expression level. To get a further insight of relative expression level ratios please refer to Fig. 1. Significant differences where only detectable for SAM towards NAM. For further statistic exploration please refer to Table 4. hADSC, human adipogenic-derived stroma cell; hMSC, human bone marrow mesenchymal stroma cell; NAM, normal adipogenic medium; SAM, specific adipogenic medium.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4488646&req=5

Figure 3: Flowchart shows the allocation of adipogenic growth media (SAM and NAM) with each cell type (hADSC and hMSC). Arrows show up- and downregulation of the relative gene expression level. To get a further insight of relative expression level ratios please refer to Fig. 1. Significant differences where only detectable for SAM towards NAM. For further statistic exploration please refer to Table 4. hADSC, human adipogenic-derived stroma cell; hMSC, human bone marrow mesenchymal stroma cell; NAM, normal adipogenic medium; SAM, specific adipogenic medium.
Mentions: At the second primer point at day 21 (t2), there were only measurable significant differences for LPL (P≤0.01) and FABP4 (P≤0.05). LPL's relative expression decreased and FABP4 stayed on a constant level. The marker genes PPARγ2 and GLUT4 showed an increased relative expression compared to primer point one (t1). In contrast C/EBPα showed a constant relative expression. For the PPARγ2, C/EBPα, and GLUT4 marker gene, there were no significant differences detectable between SAM and NAM (P>0.05) (Table 4, Figs. 2, 3).

Bottom Line: Our results revealed that only SAM was significantly (P<0.05) superior in the differentiation process in contrast to NAM for 10 days and 21 days.As well was SAM superior to differentiate the used cell lineages.The results suggest that the SAM should be established as a new standard medium for a more promising in vitro adipogenic differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral and Maxillofacial Surgery, Christian-Albrechts-University, Kiel, Germany.

ABSTRACT
To date there is no sufficient in vitro fat tissue engineering and a protocol has not been well established for this purpose. Therefore, we evaluated the in vitro influence of two different adipogenic growth media for their stimulation potential on different cell lineages to clearly define the most potent adipogenic growth media for future in vitro tissue engineering approaches. The samples for differentiation were composed of human adipogenic-derived stroma cells (hADSCs) and human bone marrow mesenchymal stroma cells (hMSCs). A normal adipogenic medium (NAM) and a specific adipogenic medium (SAM) were tested for their adipogenic stimulation potential. After 10 days and 21 days the relative gene expression was measured for the adipogenic marker genes PPARγ2, C/EBPα, FABP4, LPL, and GLUT4 detected through real time reverse transcriptase polymease chain reaction (RT-PCR). Other study variables were the comparison between NAM and SAM and between the used cells hADSCs and hMSCs. Additionally an Oil-Red staining was performed after 21 days. Our results revealed that only SAM was significantly (P<0.05) superior in the differentiation process in contrast to NAM for 10 days and 21 days. As well was SAM superior to differentiate the used cell lineages. This was evaluated by the detected marker genes PPARγ2, C/EBPα, FABP4, LPL, and GLUT4 through real time RT-PCR and by Oil-Red staining. In addition, the hMSCs proofed to be equal donor cells for adipogenic differentiation especially when stimulated by SAM. The results suggest that the SAM should be established as a new standard medium for a more promising in vitro adipogenic differentiation.

No MeSH data available.