Background and Objectives. There is a global trend towards an increase in the number of patients diagnosed with ovarian cancer during their reproductive years. One of the current clinical technologies is the technology of cryopreservation of removed healthy ovaries in order to preserve fertility and their subsequent transplantation after treatment for cancer of other organs. Glycerol is often used as a non-penetrating agent in freezing to improve follicle survival. Materials and Methods. The work examined the ovaries of cats with diagnoses confirmed by histological studies: follicular phase, luteal phase, serous carcinoma, leimyosarcoma. Diffuse reflectance spectroscopy was used to determine the kinetic parameters of dehydration and optical properties of tissues upon interaction with glycerol. Based on the change in mass over a long period of time, the diffusion coefficient of glycerol in the samples was determined. Results. The effective diffusion coefficient of interstitial water in cat ovarian tissue has been measured: D = (2.6 ± 0.4)·10⁻⁶ cm²/s (follicular phase), D = (3.3 ± 0.4)·10⁻⁶ cm²/s (luteal phase), D = (3.0 ± 0.3)·10⁻⁶ cm²/s (leiomyosarcoma), and D = (1.6 ± 0.2)·10⁻⁶ cm²/s (serous carcinoma), which is initiated within 1.5–2 hours of interaction. Diffusion of glycerol occurs over a long period of time, about 400 hours, and for the samples under study is: D = (8.3 ± 2.5)·10⁻⁸ cm²/s (follicular phase), D = (5.6 ± 1.7)·10⁻⁸ cm²/s (luteal phase), D = (2.2 ± 0.2)·10⁻⁸ cm²/s (leiomyosarcoma), and D = (1.1 ± 0.4)·10⁻⁷ cm²/s (serous carcinoma). Conclusion. The established perfusion-kinetic parameters of glycerol/interstitial water for the studied samples can be used in clinical practice in the preparation of ovarian tissue for transplantation (cryopreservation), in the transmembrane transfer of drugs, the development of new reproductive technologies, etc.