Abstract
Human pluripotent stem cells (hPSCs) are a promising source of autologous endothelial progenitor cells (EPCs) that can be used for the treatment of vascular diseases. However, this kind of treatment requires a large amount of EPCs. Therefore, a highly efficient, robust, and easily reproducible differentiation protocol is necessary. We present a novel serum-free differentiation protocol that exploits the synergy of multiple powerful differentiation effectors. Our protocol follows the proper physiological pathway by differentiating EPCs from hPSCs in three phases that mimic in vivo embryonic vascular development. Specifically, hPSCs are differentiated into (i) primitive streak, which is subsequently turned into (ii) mesoderm, which finally differentiates into (iii) EPCs. This differentiation process yields up to 15 differentiated cells per seeded hPSC in 5 days. Endothelial progenitor cells constitute up to 97% of these derived cells. The experiments were performed on the human embryonic stem cell line H9 and six human induced pluripotent stem cell lines generated in our laboratory. Therefore, robustness was verified using many hPSC lines. Two previously established protocols were also adapted and compared to our synergistic three-phase protocol. Increased efficiency and decreased variability were observed for our differentiation protocol in comparison to the other tested protocols. Furthermore, EPCs derived from hPSCs by our protocol expressed the high-proliferative-potential EPC marker CD157 on their surface in addition to the standard EPC surface markers CD31, CD144, CD34, KDR, and CXCR4. Our protocol enables efficient fully defined production of autologous endothelial progenitors for research and clinical applications.
Highlights
According to the World Health Organization, ischemic heart disease and stroke have been two major causes of death worldwide for the last 15 years
Expression of pan-endothelial markers was similar in both cell types, but expression of all the endothelial colony-forming cells (ECFCs) markers was much lower in the human saphenous vein ECs (HSVECs) than in the derived ECFCs (Figure 3B)
It was recently discovered that CD157 is present on the surface of all HPP-ECFCs, but it was missing on standard ECFCs and mature endothelial cells (ECs) in vivo (Wakabayashi et al, 2018)
Summary
According to the World Health Organization, ischemic heart disease and stroke have been two major causes of death worldwide for the last 15 years. Attempts at efficient in vitro endothelial differentiation of hPSCs have been conducted for at least 10 years (Choi et al, 2009; Park et al, 2010; Vodyanik et al, 2010; Joo et al, 2011; Li et al, 2011; Tatsumi et al, 2011; Adams et al, 2013; Prasain et al, 2014; Sahara et al, 2014; Zhang et al, 2014; Bao et al, 2015; Patsch et al, 2015; Sriram et al, 2015; Kitajima et al, 2016; Ye et al, 2016; Harding et al, 2017; Olmer et al, 2018; Suknuntha et al, 2018; Zhao et al, 2018) This strategy has the potential to ensure a consistent and unlimited source of ECFCs for in vitro studies and regenerative medicine. C2—a HSVEC line from single donor isolated in our laboratory
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
