关键词: 鸡, 原始生殖细胞, 冷冻保存, 家禽保种

Abstract: To identify the optimal cryopreservation protocol for chicken primordial germ cells（PGCs），eight cryopreservation solutions — including both literature ⁃ reported and commercially available formulations—were evaluated. Two different cell concentrations were cryopreserved under identical culture conditions and processing method. Post⁃thaw analyses，including trypan blue exclusion assay，CCK⁃8 cell proliferation assay，and apoptosis detection were used to performed to assess cell viability and status in each group，thereby evaluating the performance of the eight cryoprotectants at each cell concentration. The results showed that t Cryoprotectant 7 yielded the highest number of viable cells at a concentration of 1.37×106 cells/mL，reaching 1.14×106，with 74.8% of cell being negative for both fluorescein isothiocyanate（FITC）and propidium iodide（PI）. In contrast，Cryoprotectant 1 yielded the highest number of viable cells at 2.75×106 cells/mL，reaching 2.60×106，with 75.2% ouble⁃negative cells . However， Cryoprotectants II and Ⅷ showed the least favorable outcomes. These findings indicate that the effectiveness of cryoprotectants varies with cell concentration. The most suitable solution for cryopreserving PGCs at 1.37×106 cells/mL in 250 μL was Solution Ⅶ（Saichang Biological Solution）. For cryopreservation solution PGCs at 2.75×106 cells/mL， the optimal formulation was SolutionⅠ（basal medium∶chicken serum∶DMSO∶CaCl2=81.25%∶10%∶8%∶0.75%）.

Key words: Chicken, Primitive germ cells, Frozen preservation, Poultry preservation