The following are the supplementary data related to this art

The following are the supplementary data related to this article.
Author contribution
Acknowledgements A considerable part of the cost of this study was covered by funds of the National Strategic Reference Framework (NSRF) 2007–2013 (Region of Epirus) (NSRF/2007-13/2011ΕΠ01880025). The title of the funded proposal was “Confronting the Rupture of Anterior Cruciate Ligament: Regeneration using Stem Cells and Tissue Engineering” and T. Fotsis being the principal investigator.
Resource table
Resource details This study was approved by the Johns Hopkins University Institutional Review Board (IRB; protocol number, IRB#CR00007000), Baltimore MD. To protect patient privacy, no personal patient information is presented here. Peripheral blood (PB) was obtained from a 12-year-old boy compound heterozygous for null frameshift mutations in ATM exon 23 (3369delA in codon 1123) and exon 26 (3754delTATinsCA in codon 1252). Homozygous mutations in ATM cause the autosomal recessive syndrome Ataxia-Telangiectasia (A-T) and, accordingly, the patient clinical presentation was consistent with the “classical” form of A-T. Peripheral blood mononuclear ABT888 (PB MNC) were briefly expanded in vitro with a cytokine cocktail that stimulates erythroblast proliferation and nucleofected with three plasmid vectors encoding hOCT4, hSOX2, hC-MYC, hKLF4 and hBCL2L1, as we had described (Chou et al., 2015). Emerging TRA-1-60+ colonies were pooled and expanded. This technique successfully established feeder-free, xeno-free iPSC. Episomal plasmid integration was analyzed by qPCR on iPSC DNA at passage 22 with plasmid-specific primers, using S18 primers as loading control (Fig. 1A). These analyses confirmed lack of integration of the reprogramming plasmids. The cells were karyotypically normal (Fig. 1B) and Sanger sequencing confirmed the presence of the 3369delA in exon 23 and 3754delTATinsCA in exon 26 (Fig. 1C). Indirect immunofluorescence confirmed the expression of stem cells markers TRA-1-60, OCT4, SOX2 and SEEA4 in single cells (Fig. 1D). Finally, the SF-003 iPSC line formed teratomas in vitro with high efficiency (Fig. 1E), further confirming pluripotency.
Materials and methods
Verification and authentication For karyotyping, SF-003 cells at passage 10 were sent to WiCell Genetic Laboratory for metaphse preparation and G banding. A total of 20 metaphases were analyzed. No karyotypic abnormalities were detected. To confirm line identity, iPSC genomic DNA was sequenced using primers specific for exons 23 and 26 containing the mutations (see Table 1 for primer sequences). To assess purity, we confirmed expression of pluripotency markers TRA-1-60, OCT4, SOX4 and SSE4 in single cells by indirect immunofluorescence.
Acknowledgements
Resource table
Resource details Induced pluripotent stem cells (iPSCs) was derived from a male retinoblastoma patient after informed consent. Sequencing analysis confirmed a heterozygous missense mutation c.2663G>A (p.S888A) in the 25 exon of RB1 in RB-iPS cells (Fig. 1A). This mutation located within exon and intron junction which may cause the alternative splicing. By using primers that spans the junction of exons 23 and 26, two different size products of RB1 gene were found in RB patient-derived skin fibroblast cells (RB-SF) and RB-iPS cells, but not in the normal skin fibroblast cells (N-SF) (Fig. 1B). Similar to the human embryonic stem cell line (chHES-8) which were obtained from the established hESC bank at our center (Lin et al., 2009), the RB-iPS cells showed high telomerase activity compare to the skin fibroblasts and expressed the pluripotency related genes by RT-PCR (Fig. 1C–D). During long-term culture on the mitotically inactivated mouse embryonic fibroblasts (MEFs), the RB-iPS cells maintained a stable karyotype 46, XY (Fig. 1E), and were positive for OCT4, NANOG, SSEA4, TRA-1-60 and TRA-1-81 as well as alkaline phosphatase (Fig. 1F). The differentiation capacity of RB-iPS cells was confirmed through in vitro assays. The cells from embryoid bodies expressed the key genes related with the development of main organs from all three germ layers, such as ectoderm marker β-TUBULIN, mesoderm marker SMA and endoderm marker AFP (Fig. 1G).