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

摘要： 为寻找鸡原始生殖细胞（primordial germ cells，PGCs）的最佳冻存方案，选择近年来 文献中提到的以及市面上流通的八种冻存液，使用相同处理方法对相同培养条件的两种细胞 量进行冻存。复苏后通过台盼蓝计数法、CCK8?细胞增殖试验和细胞凋亡试验检测各组的细胞 量和细胞状态，来对八种冻存液在两个细胞量中的表现进行评价。结果表明：Ⅶ号冻存液在 1.37×106 cells/mL细胞量中的活细胞回收数最高，达到1.14×106个，且异硫氰酸荧光素（FITC） 和碘化丙啶（PI）均阴性的细胞达到74.8%；Ⅰ号冻存液在2.75×106 cells/mL细胞量中的活细胞 回收数最高，达到2.60×106个，FITC和PI均阴性的细胞达到75.2%。而Ⅱ号和Ⅷ号冻存液效果 最差。试验可以得出结论：不同的冻存液在不同细胞量中表现的冻存效果不同。在250 μL 冻 存液中，最适合1.37×106 cells/mL PGCs的是Ⅶ号冻存液（赛存生物冻存液）；最适合2.75×106 cells/mL PGCs的是Ⅰ号冻存液（基础培养基∶鸡血清∶DMSO∶CaCl2=81.25%∶10%∶8%∶0.75%）。

关键词: 鸡, 原始生殖细胞, 冷冻保存, 家禽保种