Considerable evidence suggests that breast cancer development and metastasis are driven by cancer stem-like cells (CSCs). Due to their unique role in tumor initiation, the interaction between CSCs and stromal cells is especially critical. In this work, we developed a platform to reliably isolate single cells in suspension and grow single-cell-derived spheres for functional enrichment of CSCs. The platform also allows adherent culture of stromal cells for cancer-stromal interaction. As a proof of concept, we grew SUM149 breast cancer cells and successfully formed single-cell-derived spheres. Cancer-associated fibroblasts (CAFs) as stromal cells were found to significantly enhance the formation and growth of cancer spheres, indicating elevated tumor-initiation potential. After on-chip culture for 14 days, we retrieved single-cell derived spheres with and without CAF co-culture for single-cell transcriptome sequencing. Whole transcriptome analysis highlights that CAF co-culture can boost cancer stemness especially ALDHhigh CSCs and alter epithelial/mesenchymal status. Single-cell resolution allows identification of individual CSCs and investigation of cancer cellular heterogeneity. Incorporating whole transcriptome sequencing data with public patient database, we discovered novel genes associated with cancer-CAF interaction and critical to patient survival. The preliminary works demonstrated a reliable platform for enrichment of CSCs and studies of cancer-stromal interaction.

中文翻译：

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功能丰富的癌症干细胞样细胞和癌症相关的成纤维细胞的共培养，用于单细胞全转录组分析。

大量证据表明，乳腺癌的发生和转移是由癌干样细胞（CSC）驱动的。由于它们在肿瘤引发中的独特作用，CSC与基质细胞之间的相互作用尤为关键。在这项工作中，我们开发了一个平台，该平台可可靠地分离悬浮中的单细胞并培养单细胞来源的球体，以丰富CSC的功能。该平台还允许基质细胞的贴壁培养，以实现癌症与基质之间的相互作用。作为概念验证，我们培养了SUM149乳腺癌细胞并成功地形成了源自单细胞的球体。发现与癌症相关的成纤维细胞（CAF）作为基质细胞显着增强了癌球的形成和生长，表明升高的肿瘤起始潜力。片上培养14天后，我们检索了带有和不带有CAF共培养物的单细胞衍生球体，用于单细胞转录组测序。全转录组分析表明，CAF共培养可以增强癌症的干性，尤其是ALDHhigh CSCs，并改变上皮/间质状态。单细胞分辨率可识别单个CSC并研究癌细胞的异质性。将完整的转录组测序数据与公共患者数据库相结合，我们发现了与癌症-CAF相互作用相关且对患者生存至关重要的新基因。初步工作证明了丰富的CSCs和癌症-基质相互作用研究的可靠平台。全转录组分析表明，CAF共培养可以增强癌症的干性，尤其是ALDHhigh CSCs，并改变上皮/间质状态。单细胞分辨率可识别单个CSC并研究癌细胞的异质性。将完整的转录组测序数据与公共患者数据库相结合，我们发现了与癌症-CAF相互作用相关且对患者生存至关重要的新基因。初步工作证明了丰富的CSCs和癌症-基质相互作用研究的可靠平台。全转录组分析表明，CAF共培养可以增强癌症的干性，尤其是ALDHhigh CSCs，并改变上皮/间质状态。单细胞分辨率可识别单个CSC并研究癌细胞的异质性。将完整的转录组测序数据与公共患者数据库相结合，我们发现了与癌症-CAF相互作用相关且对患者生存至关重要的新基因。初步工作证明了丰富的CSCs和癌症-基质相互作用研究的可靠平台。我们发现了与癌症-CAF相互作用相关且对患者生存至关重要的新基因。初步工作证明了丰富的CSCs和癌症-基质相互作用研究的可靠平台。我们发现了与癌症-CAF相互作用相关且对患者生存至关重要的新基因。初步工作证明了丰富的CSCs和癌症-基质相互作用研究的可靠平台。