Expression of CD105 on expanded mesenchymal stem cells does not predict their chondrogenic potential  M.A. Cleary, R. Narcisi, K. Focke, R. van der Linden, P.A.J. Brama, G.J.V.M. van Osch  Osteoarthritis and Cartilage  Volume 24, Issue 5, Pages 868-872 (May 2016) DOI: 10.1016/j.joca.2015.11.018 Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 BMSC chondrogenic potential is not linked to the percentage of CD105 expressing BSMCs. The percentage of CD105 expressing BMSCs in a total, plastic adherent BMSC population and quantification of pellet size (mm2) and area of thionine staining (%) of GAGs (Mean ± 95% CI of one donor) of the chondrogenically differentiated (3–5 weeks) total BMSC population (1–4 replicates/donor) after (a) four culture passages (N = 6 donors; 54–91 years), (b) two culture passages (N = 7 donors; 54–91 years) or (c) over multiple culture passages (Passages 1–4, N = 2 donors, 83 & 72 years; Passages 1, 2 & 5, N = 2 donors, 56 & 79 years). Passage number below histology image indicates passage number at which pellets harvested. X = no pellet formed. Images are representative images. Number on data point represents number of replicates/donor. Osteoarthritis and Cartilage 2016 24, 868-872DOI: (10.1016/j.joca.2015.11.018) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 CD105Bright and CD105Dim BMSCs show no difference in chondrogenic potential. (a) CD105 staining and sorting efficiency of a representative total, plastic adherent BMSC population (Donor1, 88 years, Passage 5), (b) cell size of CD105 subpopulations, (c) Hoechst viability staining of total, plastic adherent BMSC population before sorting, (d) cell morphology of CD105 subpopulations (Donor2, 48 years, Passage 3), (e) CD105 staining of a total, plastic adherent BMSC population from which Bright and Dim CD105 expressing BMSCs were sorted and assessed for purity, (f) quantification of thionine staining of GAGs and collagen type II staining of the chondrogenically differentiated (3–5 weeks) CD105+ subpopulations. Mean ± 95% CI of all replicates from four donors (54–79 years; Bright-Donor1: n = 1 replicate, Donor2: n = 2 replicates, Donor3: n = 1 replicate, Donor4: n = 2 replicates; Dim-Donor1: n = 2 replicates, Donor2: n = 2/3 replicates, Donor3: n = 4 replicates, Donor4: n = 2 replicates). Thionine – linear mixed model analysis (P = 0.487); Collagen type II – no statistical analysis. SSC = side scattered light, FSC = forward scattered light. Osteoarthritis and Cartilage 2016 24, 868-872DOI: (10.1016/j.joca.2015.11.018) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions