Cellular uptake of nanoliposomes with optimized cholesterol concentration and lipid extract derived from a model low grade serous ovarian carcinoma cell line

Introduction: Low-grade serous ovarian carcinoma (LGSOC) represents a rare subtype of epithelial ovarian cancer, accounting for less than 10% of all cases and typically diagnosed in younger women. The disease is characterized by tumor growth driven by activating mutations in the MAPK pathway, including KRAS , BRAF , and NRAS . Despite its slow progression, LGSOC remains challenging to treat due to limited responsiveness to conventional chemotherapy. The objective of this study was to isolate cellular lipid extract (LE) material from a model LGSOC cell line, and to utilize the newly acquired LE material to develop a relatively target specific nanoliposomal system. Early formulation and in vitro studies include optimization for cholesterol and LE content. Methods: Low-grade serous ovarian carcinoma cell line (PM-LGSOC-01, Cytion) was cultured and expanded in EMEM growth medium supplemented with 10% FBS. The PM-LGSOC-01-lipid extract (LE) material was extracted from PM-LGSOC-01 when the cells reached ~90% confluency. Lipid extraction was performed as described previously (Alharbi & Campbell, AAPS Open, number: 5(2018)). Nanoliposomal preparations consisted of DOPC, cholesterol, and LE at various ratios and wereformed by thin film method using a Buchi R-80 rotary evaporator. DPPE-Rhodamine was included in the preparations for cellular studies. Following sonication, particle size and zeta potential values weredetermined using ZetaPals. Fluorescence detection was performed using a fluorescence microplate reader. Results: Three preparations (1) DOPC (100%), (2) DOPC/Chol (95/5), and (3) DOPC/Chol (90/10) wereprepared with an average particle size of 221 ± 3 nm, 250 ± 5 nm, and 139 ± 0.3 nm, respectively. Preliminary results suggest that the additional inclusion of cholesterol in the nanoliposomal preparations increased their uptake by the target (PM-LGSOC-01) cells. Studies investigating the influence of PM-LGSOC-01-LE on the uptake of nanoliposomes by the target cells are currently underway. Conclusion: To date, the influence of cholesterol on the uptake of nanoliposomes by LGSOC cells has been investigated. Future studies will evaluate the effect of optimized cholesterol and LE contenton targeting low-grade serous ovarian carcinoma.