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Polymersomes are vesicular constructs formed by the self‐assembly of polymer amphiphiles. Here we report the fabrication of PEtOx‐b‐PLA polymersomes at physiological pH/salt concentration and the determination of a fPEtOx range, which is one of the most recognized factors that governs the self‐assembly of amphiphilic copolymers, for the first time. The findings indicate that PEtOx‐b‐PLA copolymers, containing PEtOx fractions of varied lengths, self‐assemble to form polymersomes in the range of 98.35 to 119.50 nm. The encapsulation potential of PEtOx‐b‐PLA polymersomes was demonstrated by loading of docetaxel (hydrophobic) with 17.90% encapsulation efficiency and by loading of pDNA (hydrophilic) with 65.03% encapsulation efficiency. In addition, size distribution profiles showed that polymersomes are colloidally stable for 24 h at 37 °C and for 3 months at +4 °C. The cell viability results showed that PEtOx‐b‐PLA copolymer and polymersomes are not cytotoxic to HUVEC, HEK293 and hMSC cell lines.

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