In recent years, stem cell therapy has shown promise in regenerative medicine. The lack of standardized protocols for cell isolation and differentiation generates conflicting results in this field. Mesenchymal stem cells derived from adipose tissue (ASC) and fibroblasts (FIB) share very similar cell membrane markers. In this context, the distinction of mesenchymal stem cells from fibroblasts has been crucial for safe clinical application of these cells. In the present study, we developed aptamers capable of specifically recognize ASC using the Cell-SELEX technique. We tested the affinity of ASC aptamers compared to dermal FIB. Quantitative PCR was advantageous for the in vitro validation of four candidate aptamers. The binding capabilities of Apta 2 and Apta 42 could not distinguish both cell types. At the same time, Apta 21 and Apta 99 showed a better binding capacity to ASC with dissociation constants (Kd) of 50.46 ± 2.28 nM and 72.71 ± 10.3 nM, respectively. However, Apta 21 showed a Kd of 86.78 ± 9.14 nM when incubated with FIB. Therefore, only Apta 99 showed specificity to detect ASC by total internal reflection microscopy (TIRF). This aptamer is a promising tool for the in vitro identification of ASC. These results will help understand the differences between these two cell types for more specific and precise cell therapies.

近年来，干细胞疗法在再生医学中显示出前景。缺乏用于细胞分离和分化的标准化协议在该领域产生了相互矛盾的结果。源自脂肪组织 (ASC) 和成纤维细胞 (FIB) 的间充质干细胞具有非常相似的细胞膜标记。在这种情况下，间充质干细胞与成纤维细胞的区别对于这些细胞的安全临床应用至关重要。在本研究中，我们开发了能够使用 Cell-SELEX 技术特异性识别 ASC 的适体。我们测试了 ASC 适体与真皮 FIB 相比的亲和力。定量 PCR 有利于四种候选适体的体外验证。Apta 2 和 Apta 42 的结合能力无法区分这两种细胞类型。同时，Apta 21 和 Apta 99 显示出更好的 ASC 结合能力，解离常数 (Kd) 分别为 50.46 ± 2.28 nM 和 72.71 ± 10.3 nM。然而，当与 FIB 一起孵育时，Apta 21 的 Kd 为 86.78 ± 9.14 nM。因此，只有 Apta 99 显示出通过全内反射显微镜 (TIRF) 检测 ASC 的特异性。该适体是用于体外鉴定 ASC 的有前途的工具。这些结果将有助于了解这两种细胞类型之间的差异，以进行更具体和更精确的细胞疗法。该适体是用于体外鉴定 ASC 的有前途的工具。这些结果将有助于了解这两种细胞类型之间的差异，以进行更具体和更精确的细胞疗法。该适体是用于体外鉴定 ASC 的有前途的工具。这些结果将有助于了解这两种细胞类型之间的差异，以进行更具体和更精确的细胞疗法。