Supporting Information: Virus Activated Artificial ECM Induces the Osteogenic Differentiation of Mesenchymal Stem Cells Without Osteogenic Supplements
By Jianglin Wang, Lin Wang, Xin Li, and Chuanbin Mao*
Department of Chemistry and Biochemistry, Stephenson Life Sciences Research Center, University of Oklahoma, Norman, OK 73019, USA Email:
[email protected]
Table S1. The primer sequencesfor Real-time PCR analysis Gene
Sequence of primer
OCN
5’-AAAGCCCAGCGACTCT-3’
Length of fragment 217bp
5’-CTAAACGGTGGTGCCATAGAT-3’ OPN
5'-GACGGCCGAGGTGATAGCTT-3'
209bp
5'-CATGGCTGGTCTTCCCGTTGC-3' COL
5’-TCCTGCCGATGTCGCTATC-3’
235bp
5’-CAAGTTCCGGTGTGACTGGTG-3’ Arbp
5'-CGACCTGGAAGTCCAACTAC-3' 5'-ATCTGCTGCATCTGCTTG-3'
1
109bp
Figure S1. Cell morphologies of MSCs on different substrates. Cell alignment could be obviously observed in a large field on the WT-phage film materials (A, B). There was no obvious cell alignment on the control substrate made of the polylysine without phage (C, D). Cell nuclei were stained by DAPI (blue) and F-actin were stained by FITC-labeled phalloidin (green).
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Figure S2. Immunofluorescence images of two specific osteogenic markers, osteocalcin (OCN) and osteopontin (OPN), and a non-specific osteogenic protein collagen I (COL) on different phage types (i.e., phage with different peptides displayed on the side walls) in the osteogenic differentiation media. WT, RGD, PHSRN and RGD/PHSRN denote films made of wild type phage, RGD-displayed phage, PHSRN-displayed phage, and a mixture of RGD- and PHSRN-displayed phage, respectively. CON denotes control (polylysine substrates without phage). OCN, OPN and COL were stained by rhodamine-labeled antibody (red) and cell nuclei were stained by DAPI (blue) and F-actin were stained by FITC-labeled phalloidin (green).
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Figure S3. Real-time PCR analysis of the relative expression for osteogenic genes of osteocalcin (OCN) and osteopontin (OPN), and non-specific osteogenic gene of collagen I (COL) from the cells growing on phage-based film materials in the osteogenic differentiation media. The gene expression level of both OCN and OPN was up-regulated. The group of RGD/PHSRN-phage presented the maximum mRNA expression of OCN and OPN, which was greatly higher than the control. All data represented the mean±SD (n=3, * p ＜0.05, **p＜0.01). WT, RGD, PHSRN and RGD/PHSRN denote films made of wild type phage, RGD-displayed phage, PHSRN-displayed phage, and a mixture of RGD- and PHSRNdisplayed phage, respectively.
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