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Human platelet lysate as a fetal bovine serum substitute improves human adipose-derived stromal cell culture for future cardiac repair applications.

Naaijkens BA, Niessen HW, Prins HJ, Krijnen PA, Kokhuis TJ, de Jong N, van Hinsbergh VW, Kamp O, Helder MN, Musters RJ, van Dijk A, Juffermans LJ - Cell Tissue Res. (2012)

Bottom Line: We found that PL-cultured ASC had a significant 3-fold increased proliferation rate and a significantly higher attachment to tissue culture plastic as well as to endothelial cells compared with FBS-cultured ASC.PL-cultured ASC showed a significantly higher migration rate compared with FBS-cultured ASC in a transwell assay.In conclusion, this study showed that culturing ASC is more favorable in PL-supplemented medium compared with FBS-supplemented medium.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands. b.naaijkens@vumc.nl

ABSTRACT
Adipose-derived stromal cells (ASC) are promising candidates for cell therapy, for example to treat myocardial infarction. Commonly, fetal bovine serum (FBS) is used in ASC culturing. However, FBS has several disadvantages. Its effects differ between batches and, when applied clinically, transmission of pathogens and antibody development against FBS are possible. In this study, we investigated whether FBS can be substituted by human platelet lysate (PL) in ASC culture, without affecting functional capacities particularly important for cardiac repair application of ASC. We found that PL-cultured ASC had a significant 3-fold increased proliferation rate and a significantly higher attachment to tissue culture plastic as well as to endothelial cells compared with FBS-cultured ASC. PL-cultured ASC remained a significant 25% smaller than FBS-cultured ASC. Both showed a comparable surface marker profile, with the exception of significantly higher levels of CD73, CD90, and CD166 on PL-cultured ASC. PL-cultured ASC showed a significantly higher migration rate compared with FBS-cultured ASC in a transwell assay. Finally, FBS- and PL-cultured ASC had a similar high capacity to differentiate towards cardiomyocytes. In conclusion, this study showed that culturing ASC is more favorable in PL-supplemented medium compared with FBS-supplemented medium.

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Related in: MedlinePlus

PL-cultured ASC have a higher migration rate than FBS-cultured ASC. Migration of FBS- and PL-cultured ASC towards a stimulus (+; combination of 2% FBS and 1% PL) and without a stimulus (−) were compared. Results are shown as percentage migrated ASC of total ASC applied. Bars means ± SD. FBS: n = 7; PL: n = 6; *p < 0.05
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Fig4: PL-cultured ASC have a higher migration rate than FBS-cultured ASC. Migration of FBS- and PL-cultured ASC towards a stimulus (+; combination of 2% FBS and 1% PL) and without a stimulus (−) were compared. Results are shown as percentage migrated ASC of total ASC applied. Bars means ± SD. FBS: n = 7; PL: n = 6; *p < 0.05

Mentions: Using a transwell assay, we showed that both FBS- and PL-cultured ASC were capable of migrating towards a stimulus (a combination of 2% FBS and 1% PL in DMEM). FBS-cultured ASC showed 15.1 ± 7.1% migration in the absence of the stimulus, and showed only a 0.5-fold increase in the presence of the stimulus. In contrast, PL-cultured ASC showed 5.6 ± 5.1% migration in the absence of the stimulus, but a significant 5-fold increase (p < 0.001) in the presence of the stimulus (Fig. 4). Moreover, significantly more PL-cultured ASC migrated compared with FBS-cultured ASC in the presence of the stimulus (33.2 ± 4.4 vs. 22.3 ± 8.4%, respectively; p < 0.05).Fig. 4


Human platelet lysate as a fetal bovine serum substitute improves human adipose-derived stromal cell culture for future cardiac repair applications.

Naaijkens BA, Niessen HW, Prins HJ, Krijnen PA, Kokhuis TJ, de Jong N, van Hinsbergh VW, Kamp O, Helder MN, Musters RJ, van Dijk A, Juffermans LJ - Cell Tissue Res. (2012)

PL-cultured ASC have a higher migration rate than FBS-cultured ASC. Migration of FBS- and PL-cultured ASC towards a stimulus (+; combination of 2% FBS and 1% PL) and without a stimulus (−) were compared. Results are shown as percentage migrated ASC of total ASC applied. Bars means ± SD. FBS: n = 7; PL: n = 6; *p < 0.05
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3316780&req=5

Fig4: PL-cultured ASC have a higher migration rate than FBS-cultured ASC. Migration of FBS- and PL-cultured ASC towards a stimulus (+; combination of 2% FBS and 1% PL) and without a stimulus (−) were compared. Results are shown as percentage migrated ASC of total ASC applied. Bars means ± SD. FBS: n = 7; PL: n = 6; *p < 0.05
Mentions: Using a transwell assay, we showed that both FBS- and PL-cultured ASC were capable of migrating towards a stimulus (a combination of 2% FBS and 1% PL in DMEM). FBS-cultured ASC showed 15.1 ± 7.1% migration in the absence of the stimulus, and showed only a 0.5-fold increase in the presence of the stimulus. In contrast, PL-cultured ASC showed 5.6 ± 5.1% migration in the absence of the stimulus, but a significant 5-fold increase (p < 0.001) in the presence of the stimulus (Fig. 4). Moreover, significantly more PL-cultured ASC migrated compared with FBS-cultured ASC in the presence of the stimulus (33.2 ± 4.4 vs. 22.3 ± 8.4%, respectively; p < 0.05).Fig. 4

Bottom Line: We found that PL-cultured ASC had a significant 3-fold increased proliferation rate and a significantly higher attachment to tissue culture plastic as well as to endothelial cells compared with FBS-cultured ASC.PL-cultured ASC showed a significantly higher migration rate compared with FBS-cultured ASC in a transwell assay.In conclusion, this study showed that culturing ASC is more favorable in PL-supplemented medium compared with FBS-supplemented medium.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands. b.naaijkens@vumc.nl

ABSTRACT
Adipose-derived stromal cells (ASC) are promising candidates for cell therapy, for example to treat myocardial infarction. Commonly, fetal bovine serum (FBS) is used in ASC culturing. However, FBS has several disadvantages. Its effects differ between batches and, when applied clinically, transmission of pathogens and antibody development against FBS are possible. In this study, we investigated whether FBS can be substituted by human platelet lysate (PL) in ASC culture, without affecting functional capacities particularly important for cardiac repair application of ASC. We found that PL-cultured ASC had a significant 3-fold increased proliferation rate and a significantly higher attachment to tissue culture plastic as well as to endothelial cells compared with FBS-cultured ASC. PL-cultured ASC remained a significant 25% smaller than FBS-cultured ASC. Both showed a comparable surface marker profile, with the exception of significantly higher levels of CD73, CD90, and CD166 on PL-cultured ASC. PL-cultured ASC showed a significantly higher migration rate compared with FBS-cultured ASC in a transwell assay. Finally, FBS- and PL-cultured ASC had a similar high capacity to differentiate towards cardiomyocytes. In conclusion, this study showed that culturing ASC is more favorable in PL-supplemented medium compared with FBS-supplemented medium.

Show MeSH
Related in: MedlinePlus