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. 2019 Jul 26;9:10864. doi: 10.1038/s41598-019-47391-2

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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General experimental design and IFP-MSC/BM-MSC growth kinetics, clonogenicity, immunophenotype and chondrogenic capacity. (A) General scheme depicting cell sources, stimulation and outcomes (created with BioRender). (B) Growth rate time course for representative individual CFU-Fs within IFP- and BM-MSC cultures showing colony confluency. (C) Entire culture growth kinetics of naïve IFP- and BM-MSC cultures until confluency (left panel), show comparable growth. Effect of TNFα/IFNγ (TI) or TNFα/IFNγ/CTGF (TIC) priming on IFP- and BM-MSC culture growth kinetics (middle and right panels). Both TI- and TIC- priming resulted in diminished proliferation for IFP-MSC. (D) Clonogenic capacity of naïve IFP- and BM-MSC cultures (CFU-Fs per 10³ MSC seeded). IFP-MSC generated significantly higher number of CFU-F compared to BM-MSC. (E) Chondrogenic differentiation capacity of naïve IFP- and BM-MSC. Chondro-pellet morphology (left photomicrographs), Hematoxylin and Eosin staining (H&E, middle photomicrographs), Toluidine blue staining (right photomicrographs). (F) sGAG production on day 21 of differentiation. IFP-MSC showed significantly higher sGAG production compared to BM-MSC. sGAGs production was normalized to total DNA content, comparable between IFP- and BM-MSC. All experiments (n = 3) were performed independently (3 different donors) and data are presented as scatter plots with mean ± SD. Unpaired t-test was used for statistical analysis *p < 0.05.