Laparoendoscopic single-site surgery is performed during laparoscopic donor nephrectomy (LDN) to reduce donor invasiveness. However, the procedure is difficult and does not improve cosmesis when the incision is made at the umbilicus. Therefore, we proposed a minimally invasive LDN with a Pfannenstiel incision using size-reduced trocars (mLDN) to achieve cosmesis and operability and aimed to assess its efficacy and safety. A total of 27 patients who underwent mLDN were recruited. Data on estimated blood loss, operative time, pneumoperitoneum time, warm ischemic time (WIT), complication rate, non-steroidal anti-inflammatory drugs (NSAIDs) used, and recipient serum creatinine levels were collected retrospectively. In mLDN, the Pfannenstiel position was incised to approximately 6 cm to retrieve the kidney, and three size-reduced trocars were placed in the left upper abdomen (2.5 mm and 5 mm) and umbilicus (5 mm). The median operation time and pneumoperitoneum time were 245 and 194 minutes, respectively. The median WIT was 276 seconds, and the serum creatinine levels of the recipients at seven days and one, three, six, and 12 months were significantly improved compared with baseline. No intra- and postoperative complications (Clavien-Dindo grade ≥ 2) were observed, and no patients used additional NSAIDs after the operation. The scarring in the mLDN group was unnoticeable postoperatively. mLDN can be performed safely, with high cosmesis, and with operability similar to that of conventional LDN. Although the WIT tended to be long, the function of the harvested kidney was maintained, and the use of analgesic NSAIDs was lower in this procedure. Our procedure should be considered as an option for LDN.