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Preparation of Pooled Human Platelet Lysate (pHPL) as an Efficient Supplement for Animal Serum-Free Human Stem Cell Cultures
Published on: October 30, 2009
The Use of Human Platelet Lysate (hPL) Enables 3D Cultivation of ADSCs Derived From Human Breast Tissue
Annelie Koenen1,2, Annemarie Schroeder1,3, Toralf Reimer4
1Department of Radiotherapy and Radiation Oncology, University Medical Center Rostock, Suedring 75, Rostock 18059, Germany, uni-rostock.de.
Background:
For the application of mesenchymal stem cells (MSCs) in regenerative medicine, xeno-free ex vivo cultivation is necessary. Therefore, this study aimed to investigate whether adipose-derived stem cells (ADSCs) from human breast tissue could be cultivated with human platelet lysate (hPL)-a promising alternative derived from human platelets-instead of fetal bovine serum (FBS).
Methods:
The influence of hPL within the culture medium of ADSCs from the female breast was analyzed regarding immunophenotype, adipogenic differentiation capacity, growth kinetics, and cell cycle distribution of ADSCs. Subsequently, the formation of spheres in hPL-supplemented medium was characterized in detail.
Results:
In comparison to FBS, the supplementation of medium with hPL resulted in a change in morphology, but the immunophenotype remained unchanged. Furthermore, supplementation of the medium with hPL caused an improved adipogenic differentiation capacity, increased growth, and shortened population doubling time (PDT) in ADSCs. Additionally, the cultivation of ADSCs with hPL led to spontaneous formation of spheres.
Conclusion:
hPL is not only a replacement for the ethically controversial cell culture additive FBS but also offers a xeno-free method for the cultivation of breast tissue-derived ADSCs and thus represents an important step from bench to bedside.
