Fujii S, Yoshida S, Inagaki E, Hatou S, Tsubota K, Takahashi M, Shimmura S, & Sugita S. Immunological Properties of Neural Crest Cells Derived from Human Induced Pluripotent Stem Cells. (2018). Stem Cells Devel, 28 (1):28-43. Dose: IHC; 1:200 dilution
Shibata S, Hayashi R, Okubo T, Kudo Y, Katayama T, Ishikawa Y, Toga J, Yagi E, Honma Y, Quantock AJ, Sekiguchi K, & Nishida K. Selective Laminin-Directed Differentiation of Human Induced Pluripotent Stem Cells into Distinct Ocular Lineages. (2018). Cell Rep, 25 (6):1668-1679.
Immunostaining, flow cytometry
Lovatt M, Yam GH-F, Peh GS, Colman A, Dunn NR, & Mehta JS. Directed Differentiation of Periocular Mesenchyme from Human Embryonic Stem Cells. (2018). Differentiation, 99 62-69.
Objective: Pluripotent stem cells are attractive sources of cells for regenerative medicine, because large numbers of therapeutically useful cells can be generated. However, a detailed understanding of how to differentiate clinically relevant cell types from stem cells is fundamentally required.
Summary: Identification of cells resembling periocular mesenchyme (POM) cells in the adult cornea, located in a niche between the trabecular meshwork and peripheral endothelium. The generation and expansion of POM is an important step in the generation of a number of cells types that could prove to be clinically useful for a number of diseases of the cornea.
Dose: 1:200 for flow cytometry and immunofluorescence.
Jiao J, Tian W, Qiu P, Norton EL, Wang MM, Chen YE, & Yang B. Induced Pluripotent Stem Cells with Notch1 Gene Mutation Show Impaired Differentiation into Smooth Muscle and Endothelial Cells: Implications for Bicuspid Aortic Valve-Related Aortopathy. (2018). J Thorac Cardiovasc Surg, 156 (2):515-522.
Objective: To develop an in vitro model with human-induced pluripotent stem cells (iPSCs) to evaluate the role of NOTCH1 in smooth muscle and endothelial cell differentiation.
Summary: NOTCH1 is critical in SMC and EC differentiation of iPSCs through neural crest stem cells and cardiovascular progenitor cells, respectively. NOTCH1gene mutations may potentially contribute to the development of thoracic aortic aneurysms by affecting SMC differentiation in some patients with bicuspid aortic valve-related aortopathy.
Dose: Immunofluorescence staining and flow cytometry was performed.
Hamano S, Tomokiyo A, Hasegawa D, Yoshida S, Sugii H, Mitarai H, Fujino S, Wada N, & Maeda H. Extracellular Matrix from Periodontal Ligament Cells Could Induce the Differentiation of Induced Pluripotent Stem Cells to Periodontal Ligament Stem Cell-Like Cells. (2017). Stem Cells Dev, 27 (2):100-111. Dose: Immunofluorescence staining (1:200 dilution)
Hayashi R, Ishikawa Y, Katori R, Sasamoto Y, Taniwaki Y, Takayanagi H, Tsujikawa M, Sekiguchi K, Quantock AJ, & Nishida K. Coordinated Generation of Multiple Ocular-Like Cell Lineages and Fabrication of Functional Corneal Epithelial Cell Sheets from Human Ips Cells. (2017). Nat. Protocols, 12 (4):683-696. PMID: 28253236. ICH 1:100
Jiao J, Xiong W, Wang L, Yang J, Qiu P, Hirai H, Shao L, Milewicz D, Chen YE, Yang B. (2016) Differentiation Defect in Neural Crest-Derived Smooth Muscle Cells in Patients With Aortopathy Associated With Bicuspid Aortic Valves. EBioMedicine 10:282-90. PMID: 27394642 (Targeting Trends 17q1)
Mimura S, Suga M, Okada K, Kinehara M, Nikawa H, & Furue MK. Bone Morphogenetic Protein 4 Promotes Craniofacial Neural Crest Induction from Human Pluripotent Stem Cells. (2016). Int J Dev Biol, 60 (1-3):21-28. 2016/03/05. Immunocytochemistry and flow cytometry
Naylor RW, McGhee CNJ, Cowan CA, Davidson AJ, Holm TM, & Sherwin T. Derivation of Corneal Keratocyte-Like Cells from Human Induced Pluripotent Stem Cells. (2016). PLOS ONE, 11 (10):e0165464.
Slides containing cryosections were dried overnight at 4°C and then washed twice in Tris Buffered Saline containing 0.1% Triton X100 (TBST). Slides were then placed in block solution (3% BSA, 5% Goat serum in TBST) for at least one hour. The primary antibody was then applied in the same block solution (1:100) and left overnight at 4°C.
McCabe KL, Kunzevitzky NJ, Chiswell BP, Xia X, Goldberg JL, & Lanza R. Efficient Generation of Human Embryonic Stem Cell-Derived Corneal Endothelial Cells by Directed Differentiation. (2015). PLOS ONE, 10 (12):e0145266.
For immunostaining of the expression of Zona Occludens protein 1 (ZO-1), von Willebrand factor (vWF), p75/NGFR and CD31, traditional methods were utilized. Anti-NGFr (p75) 1:100.
Werner A, Iwasaki S, McGourty CA, Medina-Ruiz S, Teerikorpi N, Fedrigo I, Ingolia NT, & Rape M. Cell-fate determination by ubiquitin-dependent regulation of translation. (2015). Nature, 525 (7570):523-527.
Dose: Immunofluorescence 1:100.
Bajo VM, Leach ND, Cordery PM, Nodal FR, King AJ. (2014) The cholinergic basal forebrain in the ferret and its inputs to the auditory cortex. Eur J
Zeltner N, Lafaille FG, Fattahi F, & Studer L. Feeder-Free Derivation of Neural Crest Progenitor Cells from Human Pluripotent Stem Cells. (2014). JoVE(87):e51609.
Kreitzer FR, Salomonis N, Sheehan A, Huang M, Park JS, Spindler MJ, Lizarraga P, Weiss WA, So P-L, & Conklin BR. A Robust Method to Derive Functional Neural Crest Cells from Human Pluripotent Stem Cells. (2013). Am J Stem Cells, 2 (2):119-131. ICC 1:200
Menendez L, Kulik MJ, Page AT, Park SS, Lauderdale JD, Cunningham ML, & Dalton S. Directed differentiation of human pluripotent cells to neural crest stem cells. (2013). Nature Protocols, 8 203.
Summary: This protocol describes in detail how to perform a highly efficient, lineage-specific differentiation of human pluripotent cells to a NCSC fate.
Dose: 0.2 ul per 106 cells
Neurosci 40(6):2922-2940. (Targeting Trends 14q3)
Ohta S, Imaizumi Y, Okada Y, Akamatsu W, Kuwahara R, Ohyama M, Amagai M, Matsuzaki Y, Yamanaka S, Okano H, & Kawakami Y. Generation of Human Melanocytes from Induced Pluripotent Stem Cells. (2011). PLOS ONE, 6 (1):e16182.
Summary: For immunocytochemistry, cells were fixed with PBS containing 4% PFA for 20 min at room temperature. Then, cells were subjected to immunofluorescence staining (1∶100).
Lee G, Chambers SM, Tomishima MJ, & Studer L. Derivation of neural crest cells from human pluripotent stem cells. (2010). Nature Protocols, 5 688.
Summary: Protocols are presented for the purification and propagation of hPSC-NC cells using flow cytometry and defined in vitro culture conditions. This protocol has been validated in multiple independent hESC and hiPSC lines. The average time required for generating purified hPSC-NC precursors using this protocol is 2–5 weeks.
Dose: Neural crest cells (1:200).
Hartig W, Varga C, Kacza J, Grosche J, Seeger J, Luiten PG, Brauer K, Harkany T (2002) In vivo labeling of rabbit cholinergic basal forebrain neurons with fluorochromated antibodies. NeuroReport 13(11):1395-1398. (Targeting Trends 03q1)
Ferreira G, Meurisse M, Tillet Y, Lévy F (2001) Distribution and Co-Localization of Choline Acetyltransferase and p75 Neurotrophin Receptors in the Sheep Basal Forebrain: Implications for the Use of a Specific Cholinergic Immunotoxin. Neurosci 104(2):419-439. (Targeting Trends 01q4)
Tremere LA, Pinaud R, Grosche J, Hartig W, Rasmusson DD (2000) Antibody for human p75 LNTR identifies cholinergic basal forebrain of non-primate species. NeuroReport 11(10):2177-2183. (Targeting Trends 01q1)
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