Hubrecht Institute-KNAW (Royal Netherlands Academy of Arts and Sciences) and University Medical Center, Utrecht, the Netherlands; Oncode Institute, Utrecht, the Netherlands.
Hubrecht Institute-KNAW (Royal Netherlands Academy of Arts and Sciences) and University Medical Center, Utrecht, the Netherlands; Oncode Institute, Utrecht, the Netherlands; Single Cell Discoveries, Utrecht, the Netherlands.
Cell. 2019 Oct 3;179(2):527-542.e19. doi: 10.1016/j.cell.2019.08.006.
Much of current molecular and cell biology research relies on the ability to purify cell types by fluorescence-activated cell sorting (FACS). FACS typically relies on the ability to label cell types of interest with antibodies or fluorescent transgenic constructs. However, antibody availability is often limited, and genetic manipulation is labor intensive or impossible in the case of primary human tissue. To date, no systematic method exists to enrich for cell types without a priori knowledge of cell-type markers. Here, we propose GateID, a computational method that combines single-cell transcriptomics with FACS index sorting to purify cell types of choice using only native cellular properties such as cell size, granularity, and mitochondrial content. We validate GateID by purifying various cell types from zebrafish kidney marrow and the human pancreas to high purity without resorting to specific antibodies or transgenes.
目前的许多分子和细胞生物学研究都依赖于通过荧光激活细胞分选(FACS)来纯化细胞类型的能力。FACS 通常依赖于用抗体或荧光转基因构建体标记感兴趣的细胞类型的能力。然而,抗体的可用性通常受到限制,并且在原发性人组织的情况下,遗传操作既费力又不可能。迄今为止,尚无系统的方法可以在没有预先了解细胞类型标记物的情况下富集细胞类型。在这里,我们提出了 GateID,这是一种计算方法,它将单细胞转录组学与 FACS 索引排序相结合,仅使用细胞大小、粒度和线粒体含量等固有细胞特性来纯化所需的细胞类型。我们通过从斑马鱼骨髓和人胰腺中纯化各种细胞类型来验证 GateID,纯度很高,而无需使用特定的抗体或转基因。