Quantifying Alkaline Phosphatase Positivity

Alkaline phosphate (ALP) staining was performed using a Fast Red substrate kit (Nichirei Biosciences Inc., Tokyo, Japan) according to the manufacturer’s protocol as described previously (Yamasaki et al. 2014 (link)). Images of the dish were taken utilizing LUMIX (Panasonic, Osaka, Japan) and assessed for positive area using ImageJ (Abramoff et al. 2004 ). The reprogramming efficiency was determined as the number of ALP-positive colonies per total number of infected cells.

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Hamada A., Akagi E., Obayashi F., Yamasaki S., Koizumi K., Ohtaka M., Nishimura K., Nakanishi M., Toratani S, & Okamoto T. (2020). Induction of Noonan syndrome-specific human-induced pluripotent stem cells under serum-, feeder-, and integration-free conditions. In Vitro Cellular & Developmental Biology. Animal, 56(10), 888-895.

Publication 2020

Fast Red substrate kit

Dish Phosphate

Corresponding Organization :

Other organizations : Hiroshima University Hospital, Hiroshima University, Tokiwa University, University of Tsukuba

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Positive area of ALP-positive colonies
Reprogramming efficiency (number of ALP-positive colonies per total number of infected cells)

control variables

None explicitly mentioned

controls

Positive control: None mentioned
Negative control: None mentioned

Annotations

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