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Stem Cells International
Volume 2017, Article ID 6909163, 14 pages
https://doi.org/10.1155/2017/6909163
Research Article

Effects of Macromolecular Crowding on Human Adipose Stem Cell Culture in Fetal Bovine Serum, Human Serum, and Defined Xeno-Free/Serum-Free Conditions

1Adult Stem Cell Group, BioMediTech, Faculty of Medicine and Life Sciences, University of Tampere, Tampere, Finland
2Science Center, Tampere University Hospital, Tampere, Finland
3NUS Tissue Engineering Program, Life Sciences Institute, National University of Singapore, Singapore 117510
4Department of Biomedical Engineering, National University of Singapore, Singapore 117575
5Department of Oral and Maxillofacial Diseases, University of Helsinki and Helsinki University Hospital, Helsinki, Finland
6Department of Biochemistry, National University of Singapore, Singapore 117575
7Institute of Chemistry and Biology, Zurich University of Applied Sciences (ZHAW), 8820 Wädenswil, Switzerland

Correspondence should be addressed to Mimmi Patrikoski; if.atu.ffats@iksokirtap.immim

Received 26 August 2016; Revised 27 January 2017; Accepted 16 February 2017; Published 30 March 2017

Academic Editor: Peter J. Quesenberry

Copyright © 2017 Mimmi Patrikoski et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Supplementary Figure 1. The multilineage differentiation and Col IV deposition of XF/SF expanded ASCs The osteogenic, adipogenic and chondrogenic differentiation capacity of XF/SF cultured ASCs was studied in standard conditions and under MMC after cell expansion in both standard conditions and under MMC. After 28 days of inductions, osteogenic differentiation was studied using Alizarin Red (AR) staining, adipogenic differentiation using Nile Red (NR) staining and chondrogenic differentiation using Alcian Blue (AB) staining. Osteogenic differentiation was poor in XF/SF conditions under MMC; however, a cell sample from one donor survived in XF/SF conditions under MMC and showed efficient osteogenic differentiation after induction in standard osteogenic media. Adipogenic differentiation was not efficient in any of the studied conditions in XF/SF media, although a few lipid droplets were observed after cell expansion in standard XF/SF conditions. Moderate chondrogenic differentiation was observed in standard conditions and, similarly to FBS and HS cultures, an altered histological architecture of the micro mass pellet was observed after MMC expansion. Enhanced Col IV deposition under MMC induction was observed also in XF/SF conditions. Scale bar 500 μm (AR, NR, Col IV); 50 μm (AB). Abbreviations: E+/−MMC, expansion under macromolecular crowding/in standard medium; D+/−MMC, differentiation under macromolecular crowding/in standard medium; AR, Alizarin Red; NR, Nile Red; AB, Alcian blue; Col IV, collagen IV. Supplementary Figure 2. Quantitative Alizarin Red staining of ASCs Osteogenic differentiation was studied in osteogenic induction and control cultures using quantitative Alizarin Red staining and quantified with cetylpyridinium chloride extraction. ASC inducted in HS media had the strongest capacity for osteogenic differentiation compared with FBS and XF/SF induction. When compared with non-induced cultures of the same treatment group, ASCs in HS media had significantly stronger capacity for osteogenic differentiation in all induction groups. In FBS media, significantly stronger osteogenic differentiation was observed after expansion in standard medium and induction in either standard or MMC culture compared with control cultures of the same treatment group. The osteogenic differentiation capacity of ASCs in XF/SF conditions under MMC was poor. Only one donor cell sample showed capacity for osteogenic differentiation after expansion under MMC. The XF/SF cells that were expanded and differentiated in standard conditions showed variable potential for osteogenic differentiation. Due to large donor variation no statistical differences could be established for XF/SF cells. ∗ indicates p<0.05. Data are presented as mean ± SD. Abbreviations: E+/−MMC, expansion under macromolecular crowding/in standard medium; D+/−MMC, differentiation under macromolecular crowding/in standard medium. Supplementary Figure 3. Quantitative Nile Red staining of ASCs The adipogenic differentiation was analyzed in adipogenic induction and control cultures using Nile Red staining and normalized to cell number. ASCs differentiated in FBS and HS media had a significantly stronger capacity for adipogenic differentiation in all induction cultures compared with control cultures of the same treatment group. XF/SF cells did not show potential for adipogenic differentiation. ∗ indicates p<0.05; ∗∗ indicates p<0.001. Data are presented as mean ± SD. Abbreviations: E+/−MMC, expansion under macromolecular crowding/in standard medium; D+/−MMC, differentiation under macromolecular crowding/in standard medium.

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