Quantifying Target Gene Expression in HLSC

RNA was extracted from HLSC differentiated in RCCS or bioscaffolds using the PureLink RNA kit (Ambion) and cDNA was prepared using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). Target gene expression was analysed by quantitative Real-time PCR (qRT-PCR) on ABI 7300 Real-time system and normalized to endogenous 18S rRNA expression (4319413E, ThermoFisher Scientific) as previously described^{17 (link)}. Universal ProbeLibrary probes were used for target gene expression analysis (Roche). Primers used for qRT-PCR are listed in Table S1.

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Famulari E.S., Navarro-Tableros V., Herrera Sanchez M.B., Bortolussi G., Gai M., Conti L., Silengo L., Tolosano E., Tetta C., Muro A.F., Camussi G., Fagoonee S, & Altruda F. (2020). Human liver stem cells express UGT1A1 and improve phenotype of immunocompromised Crigler Najjar syndrome type I mice. Scientific Reports, 10, 887.

Publication 2020

PureLink RNA kit High-Capacity cDNA Reverse Transcription Kit ABI 7300 Real-time system

18s rrna Cdna Gene expression Gene expression analysis Primers Quantitative real time pcr Rccs Reverse transcription

Corresponding Organization :

Other organizations : University of Turin, International Centre for Genetic Engineering and Biotechnology, Institute of Biostructure and Bioimaging

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Variable analysis

independent variables

Culture conditions: RCCS or bioscaffolds

dependent variables

Target gene expression

control variables

Endogenous 18S rRNA expression

controls

Positive control: None specified
Negative control: None specified

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