Centre for Ocular Regeneration, Professor Brien Holden Eye Research Centre, Champalimaud Translational Centre for Eye Research, L. V. Prasad Eye Institute, Hyderabad, Telangana, India.
Manipal Academy of Higher Education (MAHE), Manipal, Karnataka, India.
Invest Ophthalmol Vis Sci. 2023 Jul 3;64(10):12. doi: 10.1167/iovs.64.10.12.
The existing treatment options for dry eye disease (DED) due to lacrimal gland (LG) dysfunction are mainly palliative. Mesenchymal stem cells (MSCs) based therapies and 3D-LG organoids have been explored as a curative option for LG regeneration in animal models. Human LG epithelial cultures are previously established and, here, we aim to isolate and characterize the spindle-shaped cells obtained from primary human LG cultures in order to unveil its MSC property.
Normal human lacrimal glands were obtained from individuals undergoing LG debulking surgery. The conditions for human LG-MSC culture were standardized to obtain pure population of LG-MSCs at passage 3. Population doubling time (PDT), expression of phenotypic markers, tri-lineage differentiation, colony forming potential, and gene expression analysis were carried out to assess the phenotypic and genotypic characteristics compared to bone marrow-MSCs (BM-MSCs).
Our data show that these spindle-shaped cells exhibit similar phenotypic expression, colony-forming ability, and trilineage differentiation like BM-MSCs. Moreover, the gene expression also did not show any significant difference, except for increased IL1-β in LG-MSCs. The LG-MSCs do not express any lacrimal epithelial markers unlike LG tissue.
This study reveals the first-time evidence for the presence of MSC population within the human LGs, and these cells might play a role in maintaining healthy microenvironment within normal LG and repair in diseased LGs.
由于泪腺(LG)功能障碍导致的干眼症(DED)的现有治疗选择主要是姑息性的。间充质干细胞(MSCs)为基础的治疗和 3D-LG 类器官已被探索作为动物模型中 LG 再生的一种治疗选择。人类 LG 上皮细胞培养以前已经建立,在这里,我们旨在分离和鉴定从原代人 LG 培养物中获得的纺锤形细胞,以揭示其 MSC 特性。
从接受 LG 切除术的个体中获得正常的人泪腺。将人 LG-MSC 培养的条件标准化,以获得第 3 代时的纯 LG-MSC 群体。通过细胞倍增时间(PDT)、表型标志物表达、三系分化、集落形成潜力和基因表达分析,与骨髓间充质干细胞(BM-MSCs)进行比较,评估其表型和基因型特征。
我们的数据表明,这些纺锤形细胞表现出与 BM-MSCs 相似的表型表达、集落形成能力和三系分化。此外,基因表达也没有显示出任何显著差异,除了 LG-MSCs 中 IL1-β 的增加。LG-MSCs 不表达任何泪腺上皮标志物,与 LG 组织不同。
这项研究首次证明了 MSC 群体存在于人 LG 中,这些细胞可能在维持正常 LG 内的健康微环境和在患病 LG 中修复方面发挥作用。