a protocol to assess cell cycle and apoptosis in human and mouse pluripotent cells一个协议评估细胞周期和细胞凋亡在人类和小鼠多能细胞.pdfVIP

Edel et al. Cell Communication and Signaling 2011, 9:8 /content/9/1/8 METHODOLOGY Open Access A protocol to assess cell cycle and apoptosis in human and mouse pluripotent cells 1,2,3 1 1 1,4* Michael J Edel , Cristina Menchon , Jose Miguel Andres Vaquero and Juan Carlos Izpisua Belmonte Abstract Embryonic stem cells (ESC) and induced pluripotent stem cells (iPSCs) present a great opportunity to treat and model human disease as a cell replacement therapy. There is a growing pressure to understand better the signal transduction pathways regulating pluripotency and self-renewal of these special cells in order to deliver a safe and reliable cell based therapy in the near future. Many signal transduction pathways converge on two major cell functions associated with self-renewal and pluripotency: control of the cell cycle and apoptosis, although a standard method is lacking across the field. Here we present a detailed protocol to assess the cell cycle and apoptosis of ESC and iPSCs as a single reference point offering an easy to use standard approach across the field. Introduction with an average cycle lasting just twelve hours compared Human embryonic stem cells (hESC) have been derived to somatic cells [13,14]. A recent study functionally from the inner cell mass of blastocytes and can be main- demonstrated that Cyclin A regulates pluripotency but tained in vitro indefinitely with the addition of fibroblas- is redundant in fibroblasts suggesting it is a pluripotent tic growth factor 2 (FGF2) [1]. It has been shown that a associated cell cycle kinase [15]. Many of