Queensland Eye Institute, South Brisbane, Queensland, Australia; School of Biomedical Sciences, Faculty of Health, Queensland University of Technology, Brisbane, Queensland, Australia; Mater Medical Research Institute, South Brisbane, Queensland, Australia.
Mater Medical Research Institute, South Brisbane, Queensland, Australia; School of Medicine, University of Queensland, St. Lucia, Queensland, Australia.
Cytotherapy. 2014 Jan;16(1):64-73. doi: 10.1016/j.jcyt.2013.07.006. Epub 2013 Oct 1.
Mesenchymal stromal cells (MSCs) cultivated from the corneal limbus (L-MSCs) provide a potential source of cells for corneal repair. In the present study, we investigated the immunosuppressive properties of human L-MSCs and putative rabbit L-MSCs to develop an allogeneic therapy and animal model of L-MSC transplantation.
MSC-like cultures were established from the limbal stroma of human and rabbit (New Zealand white) corneas using either serum-supplemented medium or a commercial serum-free MSC medium (MesenCult-XF Culture Kit; Stem Cell Technologies, Melbourne, Australia). L-MSC phenotype was examined by flow cytometry. The immunosuppressive properties of L-MSC cultures were assessed using mixed leukocyte reactions. L-MSC cultures were also tested for their ability to support colony formation by primary limbal epithelial (LE) cells.
Human L-MSC cultures were typically CD34⁻, CD45⁻ and HLA-DR⁻ and CD73⁺, CD90⁺, CD105⁺ and HLA-ABC⁺. High levels (>80%) of CD146 expression were observed for L-MSC cultures grown in serum-supplemented medium but not cultures grown in MesenCult-XF (approximately 1%). Rabbit L-MSCs were approximately 95% positive for major histocompatibility complex class I and expressed lower levels of major histocompatibility complex class II (approximately 10%), CD45 (approximately 20%), CD105 (approximately 60%) and CD90 (<10%). Human L-MSCs and rabbit L-MSCs suppressed human T-cell proliferation by up to 75%. Conversely, L-MSCs from either species stimulated a 2-fold to 3-fold increase in LE cell colony formation.
L-MSCs display immunosuppressive qualities in addition to their established non-immunogenic profile and stimulate LE cell growth in vitro across species boundaries. These results support the potential use of allogeneic L-MSCs in the treatment of corneal disorders and suggest that the rabbit would provide a useful pre-clinical model.
从角膜缘（L-MSCs）培养的间充质基质细胞（MSCs）为角膜修复提供了潜在的细胞来源。在本研究中，我们研究了人 L-MSCs 和可能的兔 L-MSCs 的免疫抑制特性，以开发同种异体治疗和 L-MSC 移植的动物模型。
使用含有血清的培养基或商业化无血清 MSC 培养基（MesenCult-XF Culture Kit；Stem Cell Technologies，墨尔本，澳大利亚），从人及新西兰白兔眼角膜的角膜缘基质中建立 MSC 样培养物。通过流式细胞术检查 L-MSC 培养物的表型。通过混合淋巴细胞反应评估 L-MSC 培养物的免疫抑制特性。还测试了 L-MSC 培养物支持原代角膜缘上皮（LE）细胞集落形成的能力。
人 L-MSC 培养物通常为 CD34⁻、CD45⁻和 HLA-DR⁻，CD73⁺、CD90⁺、CD105⁺和 HLA-ABC⁺。在含有血清的培养基中培养的 L-MSC 培养物中观察到高水平（>80%）的 CD146 表达，但在 MesenCult-XF 中培养的培养物中观察不到（约 1%）。兔 L-MSCs 对主要组织相容性复合体 I 的表达约为 95%，对主要组织相容性复合体 II（约 10%）、CD45（约 20%）、CD105（约 60%）和 CD90（<10%）的表达水平较低。人 L-MSCs 和兔 L-MSCs 可抑制高达 75%的人 T 细胞增殖。相反，来自任何一种物种的 L-MSCs 均可刺激 LE 细胞集落形成增加 2 倍至 3 倍。
除了已建立的非免疫原性特征外，L-MSCs 还具有免疫抑制特性，并在种间边界刺激 LE 细胞生长。这些结果支持同种异体 L-MSCs 在治疗角膜疾病中的潜在用途，并表明兔子将提供有用的临床前模型。
