Comparison of fetal bovine serum and platelet-rich plasma on human lipoaspirate-derived mesenchymal stem cell proliferation
Background: Lipoaspirate-derived stem cells (LSCs) are very promising for regenerative medicine, e.g. to treat acute myocard infarction. Fetal bovine serum (FBS) is commonly used to propagate the LSCs. However, for its clinical application, FBS contains xeno-proteins that are potential to elicit immune rejection in patients. Platelet rich plasma (PRP) is one of the candidates to replace FBS. This study was aimed to compare the proliferation of LSCs cultured in 5% PRP, 10% PRP, and FBS containing medium (MesenCult®).
Methods: LSCs were cultured in 5% PRP/DMEM, 10% PRP/DMEM, and MesenCult®. After the primary culture reached its confluency, cells were harvested using TrypLE Select and seeded (around 20,000 viable cells) in new vessels in the same media. Passages were done until passage-5, with six replications. Population doubling time (PDT) of the three groups were analyzed using Kruskal Wallis test.
Results: LSCs showed different proliferation rates when cultured in 5% PRP/DMEM, 10% PRP/DMEM, and MesenCult®. PDT of the three experimental groups in passage 1-5 were significanly different (p < 0.05), with the lowest rank was cultured in medium of 10% PRP/DMEM.
Conclusion: The results suggest that 10% PRP/DMEM can be used as an alternative to replace FBS in LSC culture. (Med J Indones. 2013;22:146-51. doi: 10.13181/mji.v22i3.583)
Keywords: Fetal bovine serum, lipoaspirate, platelet rich plasma, stem cell
Pawitan JA. Prospect of adipose tissue derived mesenchymal stem cells in regenerative medicine. Cell & Tissue Transplantation & Therapy. 2009;2:7-9.
Lindroos B, Suuronen R and Miettinen S. The potential of adipose stem cells in regenerative medicine. Stem Cell Rev. 2011;7(2):269-91. http://dx.doi.org/10.1007/s12015-010-9193-7
Shah VK, Shalia KK. Stem Cell Therapy in Acute Myocardial Infarction: A Pot of Gold or Pandora’s Box. Stem Cells International. 2011;2011:1-20.
Spees JL, Gregory CA, Singh H, et al. Internalized antigens must be removed to prepare hypoimmunogenic mesenchymal stem cells for cell and gene therapy. Mol Ther. 2004;9(5):747-56. http://dx.doi.org/10.1016/j.ymthe.2004.02.012
Pawitan JA. Platelet rich plasma in xeno-free stem cell culture: the impact of platelet count and processing method. Curr Stem Cell Res Ther. 2012;7(5):329-35. http://dx.doi.org/10.2174/157488812802481508
Rauch C, Feifel E, Amann EM, et al. Alternatives to the use of fetal bovine serum: human platelet lysates as a serum substitute in cell culture media. ALTEX. 2011;28(4): 305-16
Pawitan JA, Liem IK, Suryani D, Bustami A, Purwoko RY. Simple lipoaspirate washing using a coffee filter. Asian Biomedicine. 2013;7(3): 333-8.
Mitchell JB, McIntosh K, Zvonic S, et al. Immunophenotype of human adipose-derived cells: temporal changes in stromal-associated and stem cell-associated markers. Stem Cells. 2006;24(2):376-85. http://dx.doi.org/10.1634/stemcells.2005-0234
Pallant J. SPSS Survival Manual: a step by step guide to data analysis using SPSS. 2nd ed. Australia: Allen & Unwin;2005.
Kakudo N, Minakata T, Mitsui T, Kushida S, Notodihardjo FZ, Kusumoto K. Proliferation-promoting effect of platelet-rich plasma on human adipose-derived stem cells and human dermal fibroblasts. Plast Reconstr Surg. 2008;122(5):1352-60. http://dx.doi.org/10.1097/PRS.0b013e3181882046
Kocaoemer A, Kern S, Kluter H, Bieback K. Human AB serum and thrombin-activated platelet-rich plasma are suitable alternatives to fetal calf serum for the expansion of mesenchymal stem cells from adipose tissue. Stem Cells. 2007;25(5):1270-78. http://dx.doi.org/10.1634/stemcells.2006-0627.
Bieback K, Hecker A, Kocaömer A, et al. Human alternatives to fetal bovine serum for the expansion of mesenchymal stromal cells from bone marrow. Stem Cells. 2009;27(9):2331-41. http://dx.doi.org/10.1002/stem.139
Korzyńska A, Zychowicz A. A method of estimation of the cell doubling time on basis of the cell culture monitoring data. Biocybernetics and Biomedical Engineering. 2008; 28(4):75-82.
Dominici M, Le Blanc K, Mueller I, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006;8(4):315-7. http://dx.doi.org/10.1080/14653240600855905
Boquest AC, Shahdadfar A, Frønsdal K, et al. Isolation and transcription profiling of purified uncultured human stromal stem cells: alteration of gene expression after in vitro cell culture. Mol Biol Cell. 2005;16(3):1131-41. http://dx.doi.org/10.1091/mbc.E04-10-0949
Izadpanah R, Kaushal D, Kriedt C, et al. Long-term in vitro expansion alters the biology of adult mesenchymal stem cells. Cancer Res. 2008;68(11):4229-38. http://dx.doi.org/10.1158/0008-5472.CAN-07-5272
Copyright (c) 2013 Des Suryani, Jeanne A. Pawitan, Jinia Lilianty, Reza Y. Purwoko, Isabella K. Liem, Lia Damayanti
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Authors who publish with Medical Journal of Indonesia agree to the following terms:
- Authors retain copyright and grant Medical Journal of Indonesia right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial License that allows others to remix, adapt, build upon the work non-commercially with an acknowledgment of the work’s authorship and initial publication in Medical Journal of Indonesia.
- Authors are permitted to copy and redistribute the journal's published version of the work non-commercially (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in Medical Journal of Indonesia.