Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, 235 Hungerford Hill Road, Ithaca, NY, 14853, USA.
Elanco Animal Health, 2500 Innovation Way, Indianapolis, IN, 46241, USA.
Stem Cell Res Ther. 2024 Nov 14;15(1):429. doi: 10.1186/s13287-024-04019-1.
The secretome of primary bovine mammosphere-derived epithelial cells (MDECs) has been shown to exert antimicrobial, regenerative, and immunomodulatory properties in vitro, which warrants its study as a potential biologic treatment with the potential to be translated to human medicine. Currently, the use of the MDEC secretome as a therapy is constrained by the limited life span of primary cell cultures and the decrease of secretome potency over cell passages.
To address these limitations, early-passage bovine MDECs were immortalized using hTERT, a human telomerase reverse transcriptase. The primary and immortal MDECs were compared morphologically, transcriptomically, and phenotypically. The functional properties and proteomic profiles of the secretome of both cell lines were evaluated and compared. All experiments were performed with both low and high passage cell cultures.
We confirmed through in vitro experiments that the secretome of immortalized MDECs, unlike that of primary cells, maintained antimicrobial and pro-migratory properties over passages, while pro-angiogenic effects of the secretome from both primary and immortalized MDECs were lost when the cells reached high passage. The secretome from primary and immortalized MDECs, at low and high passages exerted immunomodulatory effects on neutrophils in vitro.
High passage immortalized MDECs retain a bioactive secretome with antimicrobial, regenerative, and immunomodulatory properties, suggesting they may serve as a consistent cell source for therapeutic use.
原代牛乳腺小球源上皮细胞（MDECs）的分泌物已被证明具有体外的抗菌、再生和免疫调节特性，这使得它有作为一种潜在的生物学治疗方法的研究价值，有可能转化为人类医学。目前，由于原代细胞培养的寿命有限以及细胞传代过程中分泌物效力的下降，MDEC 分泌物的使用受到限制。
为了解决这些限制，使用人端粒酶逆转录酶（hTERT）对早期传代的牛 MDECs 进行永生化。比较了原代和永生化 MDECs 的形态、转录组和表型。评估并比较了这两种细胞系的分泌物的功能特性和蛋白质组谱。所有实验均在低传代和高传代细胞培养物中进行。
我们通过体外实验证实，与原代细胞不同，永生化 MDEC 的分泌物在传代过程中保持了抗菌和促迁移特性，而原代和永生化 MDEC 分泌物的促血管生成作用在细胞达到高传代时丧失。低传代和高传代的原代和永生化 MDEC 的分泌物在体外对中性粒细胞具有免疫调节作用。
高传代永生化 MDECs 保留了具有抗菌、再生和免疫调节特性的生物活性分泌产物，表明它们可能作为一种稳定的治疗用途的细胞来源。
