Department of Comprehensive Dentistry, University of Texas Health Science Center at San Antonio, San Antonio, TX, 78229-3900, USA.
Department of Biomedical Engineering, University of Texas at San Antonio, San Antonio, TX, 78249, USA.
Stem Cell Res Ther. 2022 Jul 15;13(1):306. doi: 10.1186/s13287-022-02993-y.
Current treatments for salivary gland (SG) hypofunction are palliative and do not address the underlying cause or progression of the disease. SG-derived stem cells have the potential to treat SG hypofunction, but their isolation is challenging, especially when the tissue has been damaged by disease or irradiation for head and neck cancer. In the current study, we test the hypothesis that multipotent bone marrow-derived mesenchymal stem cells (BM-MSCs) in a rat model are capable of trans-differentiating to the SG epithelial cell lineage when induced by a native SG-specific extracellular matrix (SG-ECM) and thus may be a viable substitute for repairing damaged SGs.
Rat BM-MSCs were treated with homogenates of decellularized rat SG-ECM for one hour in cell suspension and then cultured in tissue culture plates for 7 days in growth media. By day 7, the cultures contained cell aggregates and a cell monolayer. The cell aggregates were hand-selected under a dissecting microscope, transferred to a new tissue culture dish, and cultured for an additional 7 days in epithelial cell differentiation media. Cell aggregates and cells isolated from the monolayer were evaluated for expression of SG progenitor and epithelial cell specific markers, cell morphology and ultrastructure, and ability to form SG-like organoids in vivo.
The results showed that this approach was very effective and guided the trans-differentiation of a subpopulation of CD133-positive BM-MSCs to the SG epithelial cell lineage. These cells expressed amylase, tight junction proteins (Cldn 3 and 10), and markers for SG acinar (Aqp5 and Mist 1) and ductal (Krt 14) cells at both the transcript and protein levels, produced intracellular secretory granules which were morphologically identical to those found in submandibular gland, and formed SG-like organoids when implanted in the renal capsule in vivo.
The results of this study suggest the feasibility of using autologous BM-MSCs as an abundant source of stem cells for treating SG hypofunction and restoring the production of saliva in these patients.
目前治疗唾液腺（SG）功能低下的方法是姑息性的，不能解决疾病的根本原因或进展。SG 来源的干细胞具有治疗 SG 功能低下的潜力，但它们的分离具有挑战性，尤其是当组织因头颈部癌症而受到疾病或放射治疗的损害时。在目前的研究中，我们测试了这样一个假设，即在大鼠模型中，多能骨髓间充质干细胞（BM-MSCs）在受到天然 SG 特异性细胞外基质（SG-ECM）诱导时能够向 SG 上皮细胞谱系转分化，因此可能是修复受损 SG 的可行替代物。
将大鼠 BM-MSCs 用脱细胞大鼠 SG-ECM 匀浆在细胞悬浮液中处理 1 小时，然后在组织培养板中在生长培养基中培养 7 天。第 7 天，培养物中含有细胞聚集体和单层细胞。在解剖显微镜下手动选择细胞聚集体，将其转移到新的组织培养皿中，并在上皮细胞分化培养基中再培养 7 天。评估细胞聚集体和从单层细胞中分离的细胞的 SG 祖细胞和上皮细胞特异性标志物表达、细胞形态和超微结构以及在体内形成 SG 样类器官的能力。
结果表明，这种方法非常有效，指导了 CD133 阳性 BM-MSCs 的亚群向 SG 上皮细胞谱系的转分化。这些细胞在转录和蛋白质水平上表达淀粉酶、紧密连接蛋白（Cldn3 和 10）以及 SG 腺泡（Aqp5 和 Mist1）和导管（Krt14）细胞的标志物，产生形态上与颌下腺相同的细胞内分泌颗粒，并在体内植入肾囊时形成 SG 样类器官。
这项研究的结果表明，使用自体 BM-MSCs 作为治疗 SG 功能低下和恢复这些患者唾液分泌的丰富干细胞来源是可行的。
Zotero 插件
