■肾母细胞瘤(WT),或者肾母细胞瘤,是最常见的小儿肾癌.大多数WT显示出“有利的”三相组织学,其中肿瘤由胚乳组成,基质,和上皮细胞类型。新辅助化疗或弥漫性间变后的肺瘤占优势(“不利”组织学;5-8%)预示着预后较差。母细胞瘤可能提供推定的癌症干细胞(CSC),保留了肾单位祖细胞(NPCs)的分子和组织学特征,在WT内。NPC出现在后肾间质(MM)中,并在发育中的肾脏中填充帽间间质(CM)。WT胚细胞,像NPC一样,类似地表达标记,SIX2和CITED1。肿瘤异种移植是目前用于研究或治疗筛选的唯一可靠的增殖肿瘤组织的方法。因为将肿瘤作为单层体外培养的努力总是失败的。因此,迫切需要快速有效地繁殖WT干细胞以实现高通量,实时药物筛选。
■以前,我们的实验室开发了支持小鼠NPCs在培养中繁殖的生态位条件。将类似的条件应用于WT,我们评估了我们维持关键NPC“干性”标记的能力,SIX2、NCAM、和YAP1,和CSC标记ALDHI在细胞中来自五个不同的未治疗的患者肿瘤。
■因此,我们的培养条件通过快速分裂的细胞的多次传代维持了这些标记在培养的WT细胞中的表达。
■这些发现表明,我们的培养条件维持了WT卵裂虫种群,如前面所示的正常NPC。因此,我们开发了新的WT细胞系和多代体外模型,用于研究WT中的囊胚谱系/CSC。此外,该系统支持异质WT细胞的生长,可以对潜在的药物疗法进行疗效和耐药性测试。
UNASSIGNED: Wilms Tumor (WT), or nephroblastoma, is the most common pediatric kidney cancer. Most WTs display a \"favorable\" triphasic histology, in which the tumor is comprised of blastemal, stromal, and epithelial cell types. Blastemal predominance after neoadjuvant chemotherapy or diffuse anaplasia (\"unfavorable\" histology; 5-8%) portend a worse prognosis. Blastema likely provide the putative cancer stem cells (CSCs), which retain molecular and histologic features characteristic of nephron progenitor cells (NPCs), within WTs. NPCs arise in the metanephric mesenchyme (MM) and populate the cap mesenchyme (CM) in the developing kidney. WT blastemal cells, like NPCs, similarly express markers, SIX2 and CITED1. Tumor xenotransplantation is currently the only dependable method to propagate tumor tissue for research or therapeutic screening, since efforts to culture tumors in vitro as monolayers have invariably failed. Therefore, a critical need exists to propagate WT stem cells rapidly and efficiently for high-throughput, real-time drug screening.
UNASSIGNED: Previously, our lab developed niche conditions that support the propagation of murine NPCs in culture. Applying similar conditions to WTs, we assessed our ability to maintain key NPC \"stemness\" markers, SIX2, NCAM, and YAP1, and CSC marker ALDHI in cells from five distinct untreated patient tumors.
UNASSIGNED: Accordingly, our culture conditions maintained the expression of these markers in cultured WT cells through multiple passages of rapidly dividing cells.
UNASSIGNED: These findings suggest that our culture conditions sustain the WT blastemal population, as previously shown for normal NPCs. As a result, we have developed new WT cell lines and a multi-passage in vitro model for studying the blastemal lineage/CSCs in WTs. Furthermore, this system supports growth of heterogeneous WT cells, upon which potential drug therapies could be tested for efficacy and resistance.