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  • Enhanced conversion of induced neuronal cells (iN cells) from human fibroblasts: Utility in uncovering cellular deficits in mental illness-associated chromosomal abnormalities.

Enhanced conversion of induced neuronal cells (iN cells) from human fibroblasts: Utility in uncovering cellular deficits in mental illness-associated chromosomal abnormalities.

Neuroscience research (2015-08-12)
Eleonora Passeri, Ashley M Wilson, Amedeo Primerano, Mari A Kondo, Srona Sengupta, Rupali Srivastava, Minori Koga, Cassandra Obie, Peter P Zandi, Fernando S Goes, David Valle, Judith L Rapoport, Akira Sawa, Shin-ichi Kano, Koko Ishizuka
ZUSAMMENFASSUNG

The novel technology of induced neuronal cells (iN cells) is promising for translational neuroscience, as it allows the conversion of human fibroblasts into cells with postmitotic neuronal traits. However, a major technical barrier is the low conversion rate. To overcome this problem, we optimized the conversion media. Using our improved formulation, we studied how major mental illness-associated chromosomal abnormalities may impact the characteristics of iN cells. We demonstrated that our new iN cell culture protocol enabled us to obtain more precise measurement of neuronal cellular phenotypes than previous iN cell methods. Thus, this iN cell culture provides a platform to efficiently obtain possible cellular phenotypes caused by genetic differences, which can be more thoroughly studied in research using other human cell models such as induced pluripotent stem cells.

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Sigma-Aldrich
N6,2′-O-Dibutyryladenosin 3′,5′-zyklisches Monophosphat Natriumsalz Monohydrat, ≥96% (HPLC), powder
Sigma-Aldrich
Valproinsäure Natriumsalz, 98%
Sigma-Aldrich
SB 431542 Hydrat, ≥98% (HPLC), powder
Sigma-Aldrich
5-Azacytidin, ≥98% (HPLC)
Sigma-Aldrich
Ziegenserum aus kontrollierter Spenderherde, USA origin, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Monoklonaler Anti-MAP2(2a+2b)-Antikörper der Maus in Maus hergestellte Antikörper, ~2 mg/mL, clone AP-20, purified from hybridoma cell culture