iPSC DIFFERENTIATION SERVICES FOR RESEARCHERS  

Figure 1. Human iPSC-motor neuron differentiation. (A) Phase contrast image of iPSC before differentiation. Scale bar: 500 µm; (B) Neural spheres upon neural induction on Day 11. Scale bar: 200 µm;(C) Neural rosette formation in neural progenitor cells (NPCs). Scale bar: 100 µm; (D and E) The cryopreserved iPSC-derived motor neurons were recovered and cultured 200,000 viable cells in Motor Neuron Maintenance Medium on PDL / Cultrex-coated wells in the 48-well plate;  (D) The cells were fixed and stained with anti-HB9 and anti-TUJ1 on Day 2 post thaw; (E) The cells were fixed and stained with anti-MAP2 and anti-ChAT on Day 7 post thaw. Scale bar: 100 µm.

Figure 2. Characterization of Human iPSC-motor neurons. (A and B) The iPSC-motor neurons were recovered and cultured in Motor Neuron Maintenance Medium for 2 days on Cultrex-coated plates, and then the cells were collected for FACS analysis with antibodies against TUJ1 and HB9; (C-E) The iPSC-motor neurons were recovered and cultured in Motor Neuron Maintenance Medium for 7 days on Cultrex-coated plates, and then the cells were collected for FACS analysis with antibodies against FOXP1, ISL1 and NeuN; (F) iPSC-MNs cell pellets were used for mycoplasma screening by PCR amplification of a ~270 bp sequence conserved in most of the mycoplasma species and a ~617 bp sequence conserved in most cells. Positive Ctl: DNA from a tested mycoplasma positive sample; Negative Ctl 1: Lysis buffer which was used to extract gDNA; Negative Ctl 2: DNA from a tested mycoplasma negative sample.

• Liu B, Li M, Zhang L, Chen Z, Lu P. (2022). Motor neuron replacement therapy for amyotrophic lateral sclerosis. Neural Regen Res;17:1633-9 -- Learn More

• Liu, Y., Dodart, J., Tran, H., Berkovitch, S., Braun, M., Byrne, M., . . . Brown, R. H. (2021). Variant-selective stereopure oligonucleotides protect against pathologies associated with c9orf72-repeat expansion in preclinical models. Nature Communications, 12(1). doi:10.1038/s41467-021-21112-8 -- Learn More

• Osaki, T., Uzel, S. G., & Kamm, R. D. (2020). On-chip 3d neuromuscular model for drug screening and precision medicine in neuromuscular disease. Nature Protocols, 15(2), 421-449. doi:10.1038/s41596-019-0248-1 -- Learn More

• Shen, X., Beasley, S., Putman, J. N., Li, Y., Prakash, T. P., Rigo, F., . . . Corey, D. R. (2019). Efficient electroporation of neuronal cells using synthetic oligonucleotides: Identifying duplex RNA and antisense oligonucleotide activators of Human frataxin expression. RNA, 25(9), 1118-1129. doi:10.1261/rna.071290.119 -- Learn More

• Gasset-Rosa, F., Lu, S., Yu, H., Chen, C., Melamed, Z., Guo, L., . . . Cleveland, D. W. (2019). Cytoplasmic TDP-43 de-mixing independent of stress granules drives inhibition of nuclear import, loss of nuclear TDP-43, and cell death. Neuron, 102(2). doi:10.1016/j.neuron.2019.02.038 -- Learn More

• Martier, R., Liefhebber, J. M., García-Osta, A., Miniarikova, J., Cuadrado-Tejedor, M., Espelosin, M., . . . Konstantinova, P. (2019). Targeting rna-mediated toxicity in c9orf72 als and/or ftd by rnai-based gene therapy. Molecular Therapy - Nucleic Acids, 16, 26-37. doi:10.1016/j.omtn.2019.02.001 -- Learn More

• Melamed, Z., López-Erauskin, J., Baughn, M. W., Zhang, O., Drenner, K., Lin, N., Wu, D., . . . Cleveland, D. W. (2019). Premature polyadenylation-mediated loss of stathmin-2 is a hallmark of tdp-43-dependent neurodegeneration. Nature Neuroscience, 22(2), 180-190. doi:10.1038/s41593-018-0293-z -- Learn More

• Marei, H. E., Althani, A., Lashen, S., Cenciarelli, C., & Hasan, A. (2017). Genetically unmatched human Ipsc and Esc Exhibit Equivalent gene expression and neuronal differentiation potential. Scientific Reports, 7(1). doi:10.1038/s41598-017-17882-1 -- Learn More

• Osaki, T., Uzel, S. G., & Kamm, R. D. (2018). Microphysiological 3D model of amyotrophic lateral sclerosis (ALS) from human iPS-derived muscle cells and optogenetic motor neurons. Science Advances, 4(10). doi:10.1126/sciadv.aat5847 -- Learn More

• Danziger, S. A., Miller, L. R., Singh, K., Whitney, G. A., Peskind, E. R., Li, G., . . . Smith, J. J. (2017). An indicator cell assay for blood-based diagnostics. PLOS ONE, 12(6). doi:10.1371/journal.pone.0178608 -- Learn More