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. 2020 Mar 5;11:105. doi: 10.1186/s13287-020-01619-5

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — Functional analysis of subpopulations in MSCs and iMSCs. a Schematic presentation of limiting dilution assays. Either primary MSCs or iMSCs of two corresponding donors were used. Cells were seeded on four 96-well plates at decreasing concentrations (30, 10, 3, and 1 cells/well). After 15 days of incubation, wells were scored for at least 50% confluency to determine CFU-f frequencies. The same plates were then induced towards adipogenic and osteogenic lineage to compare in vitro differentiation potential. b Analysis of CFU-f frequencies of MSCs and iMSCs (N = 2; n = 4). c Representative pictures of subclones without and with adipogenic differentiation potential (BODIPY/Hoechst staining 14 days after induction of differentiation; donor 3). d For individual MSC and iMSC subclones, the percentage of cells with fat droplet formation upon adipogenic differentiation was analyzed. Due to different morphology of fat droplets the results were not directly compared in one diagram. e Representative pictures of subclones without and with osteogenic differentiation potential (Alizarin Red staining 18 days after induction of osteogenic differentiation; donor 3). f Osteogenic differentiation was quantified for individual subclones in MSCs and iMSCs. Alizarin Red staining was measured by background normalized absorption