Although incorporating proteases into sperm medium is considered the most effective procedure to eliminate camel semen viscosity, it drastically affects viability, morpho‐functional properties and, hence, fertilization potential of spermatozoa. The present work aimed at evaluating adequacy of employing magnetic nanoparticles‐based sperm purification technique for eluting impaired and apoptotic camel spermatozoa from cryopreserved semen doses following protease‐based semen liquefaction. Thirty cryopreserved semen doses (50 x 10⁶ sperm/straw) representing the following liquefaction treatments: control (untreated), 0.1 mg/ml papain or 5 U/ml bromelain were used (n = 10 straws per treatment). Immediately after thawing (38°C for 40 s), sperm concentration of each straw within treatment was readjusted to 15 x 10⁶ sperm/mL by dilution in PBS (37°C). Sperm physical and cytological properties were then assessed (non‐purified semen). Thereafter, each specimen was subjected to lectin‐functionalized DNA‐defrag magnetic nanoparticles sperm purification, and the same sperm traits were re‐evaluated after undergoing purification (purified semen). Sperm DNA fragmentation level within each group, prior to and after magnetic nano‐purification, was also determined by fluorescent imaging. The results showed a dramatic improvement (p < .05) in post‐thaw motility (%), viability (%), normal sperm (%), intact acrosome (%) and HOST‐reacted (%) spermatozoa in protease‐liquefied semen following sperm magnetic nano‐purification. Additionally, the highest (p < .05) DNA fragmentation level was recorded in all cryopreserved semen groups prior to purification, whereas the lowest (p < .05) was observed in the protease‐treated specimens after magnetic nano‐purification. These results indicate that protease‐based semen liquefaction prior to cryopreservation in conjunction with magnetic nano‐purification post‐thawing holds potential for reducing the proportion of damaged and dead spermatozoa, hence improving camel sperm fertilization competence.

中文翻译：

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凝集素功能化的DNA碎片磁性纳米粒子纯化基于蛋白酶的精液后冷冻保存的骆驼精子

尽管将蛋白酶掺入精子培养基中被认为是消除骆驼精液粘度的最有效方法，但它会严重影响精子的活力，形态功能特性，进而影响精子的受精能力。本工作旨在评估采用基于磁性纳米粒子的精子纯化技术从基于蛋白酶的精液液化后冷冻保存的精液中洗脱受损和凋亡的骆驼精子的适当性。三十种冷冻保存的精液剂量（50 x 10 ⁶精子/秸秆）代表以下液化处理：对照（​​未处理），0.1 mg / ml木瓜蛋白酶或5 U / ml菠萝蛋白酶（n =每次处理10根吸管）。解冻后（38°C持续40 s），通过在PBS（37°C）中稀释，将处理中每根稻草的精子浓度重新调整为15 x 10 ⁶精子/ mL。然后评估精子的物理和细胞学特性（未纯化的精液）。之后，对每个标本进行凝集素功能化的DNA碎片磁性纳米颗粒精子纯化，并在纯化后对相同的精子性状进行重新评估（精液）。还通过荧光成像确定了磁性纳米纯化之前和之后每组中的精子DNA片段化水平。结果显示出显着改善（p < 精子磁性纳米纯化后，蛋白酶液化精子中解冻后的活力（％），活力（％），正常精子（％），完整的顶体（％）和经HOST反应（％）的精子。此外，在 纯化之前，所有冷冻保存的精液组均记录到最高的（p <.05）DNA片段水平，而在磁性纳米纯化后的蛋白酶处理标本中观察到的最低（p <  .05）。这些结果表明，冷冻保存前基于蛋白酶的精液液化和融化后的磁性纳米纯化技术有潜力减少受损和死亡的精子的比例，从而提高骆驼精子的受精能力。