Department of Morphology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil,
Stem Cell Rev Rep. 2014 Oct;10(5):697-711. doi: 10.1007/s12015-014-9529-9.
Human orbital fat tissues are a potential source to isolate stem cells for the development of regenerative medicine therapies. For future safe clinical application of these cells, it is critical to establish animal component-free culture conditions as well as to clearly define the stem cell population characteristics differentiating them from other cell types, such as fibroblasts. Therefore, the present study aimed to compare phenotypic and functional characteristics of orbital fat-derived stem cells (OFSCs) and fibroblasts resident in the eyelid skin in donor-matched samples grown in culture medium supplemented with pooled allogeneic human serum (HS) replacing fetal bovine serum (FBS). We first investigated the proliferative effects of OFSCs on HS, and then we compared the alkaline phosphatase (AP) expression and activity, immunophenotypic profile, and in vitro multilineage differentiation potential of OFSCs side-by-side with fibroblasts. The results showed that HS enhanced OFSCs proliferation without compromising their immunophenotype, AP activity, and osteogenic, adipogenic, and chondrogenic differentiation capacities. In contrast to OFSCs, the fibroblasts did not exhibit AP expression and activity and did not have multilineage differentiation potential. The results enabled us to successfully distinguish OFSCs from fibroblasts populations, suggesting that AP expression/activity and multilineage differentiation assays can be used reliably to discriminate mesenchymal stem cells from fibroblasts. Our findings also support the feasibility of pooled allogeneic HS as a safer and more effective alternative to FBS for clinical applications.
人类眼眶脂肪组织是一种潜在的来源，可以分离干细胞，用于开发再生医学疗法。为了将来这些细胞能够安全地应用于临床，建立无动物成分的培养条件以及明确区分干细胞群体特征，将其与成纤维细胞等其他细胞类型区分开来至关重要。因此，本研究旨在比较来源于眼眶脂肪的干细胞（OFSCs）和眼脸皮肤固有成纤维细胞在补充有混合异体人血清（HS）的培养基中培养时的表型和功能特征，以替代胎牛血清（FBS）。我们首先研究了 OFSCs 对 HS 的增殖作用，然后比较了 OFSCs 和成纤维细胞的碱性磷酸酶（AP）表达和活性、免疫表型特征以及体外多向分化潜能。结果表明，HS 增强了 OFSCs 的增殖能力，而不影响其免疫表型、AP 活性以及成骨、成脂和成软骨分化能力。与 OFSCs 相反，成纤维细胞不表达 AP 且没有活性，也没有多向分化潜能。这些结果使我们能够成功区分 OFSCs 和成纤维细胞群体，表明 AP 表达/活性和多向分化测定可可靠地区分间充质干细胞和成纤维细胞。我们的研究结果还支持混合异体 HS 作为 FBS 的更安全、更有效的替代物，用于临床应用的可行性。
