Song Yang, Soto Jennifer, Lin Xiao, Hoffman Tyler, Hu Erin, Zhu Ninghao, Zarubova Jana, Wu Yifan, Tian Jing, Wong Pak Kin, Li Song
Department of Bioengineering, University of California, Los Angeles, CA 90095, USA.
Institute of Biomedical Engineering, West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu 610041, China.
Adv Funct Mater. 2025 Jan 2;35(1). doi: 10.1002/adfm.202410910. Epub 2024 Nov 30.
Cell reprogramming and manufacturing have broad applications in tissue regeneration and disease treatment. However, many derived cell types lack unique cell surface markers for protein-based cell sorting, making it difficult to isolate these cells from mixed populations. Additionally, there is a need to identify and isolate cells of interest at the early stages of cell expansion. To address this challenge, we engineered a nucleic acid-based gold nanorod (NAGNR) fluorescent biosensor that can detect the mRNA expression of intracellular markers for cell sorting. We demonstrated its application in isolating induced neuronal (iN) cells from dermal fibroblast populations during the early stages of cell reprogramming. Cell sorting based on the mRNA of the neuronal transcriptional factor Ascl1 resulted in an enrichment of iN cells from 3% to 72%, and additional sorting with the transcriptional factor Scn2 further increased iN enrichment. Moreover, NAGNR biosensors can be used in conjunction with protein marker-based cell sorting. NAGNR-sorted iN cells show a functional response to electrical stimulation in a co-culture of iN cells and muscle cells. These findings demonstrate that NAGNR-based cell sorting offers great potential for cell identification and isolation at an early stage of cell reprogramming and manufacturing.
细胞重编程和制造在组织再生和疾病治疗中具有广泛应用。然而，许多衍生细胞类型缺乏用于基于蛋白质的细胞分选的独特细胞表面标志物，这使得从混合群体中分离这些细胞变得困难。此外，需要在细胞扩增的早期阶段识别和分离感兴趣的细胞。为应对这一挑战，我们设计了一种基于核酸的金纳米棒（NAGNR）荧光生物传感器，它可以检测用于细胞分选的细胞内标志物的mRNA表达。我们展示了其在细胞重编程早期从真皮成纤维细胞群体中分离诱导神经元（iN）细胞的应用。基于神经元转录因子Ascl1的mRNA进行细胞分选，使得iN细胞的富集率从3%提高到72%，使用转录因子Scn2进行进一步分选则进一步提高了iN细胞的富集率。此外，NAGNR生物传感器可与基于蛋白质标志物的细胞分选结合使用。经NAGNR分选的iN细胞在iN细胞与肌肉细胞的共培养中对电刺激表现出功能性反应。这些发现表明，基于NAGNR的细胞分选在细胞重编程和制造的早期阶段进行细胞识别和分离方面具有巨大潜力。
