Fibrolamellar carcinomas (FLCs), lethal tumors occurring in children to young adults, have genetic signatures implicating derivation from biliary tree stem cell (BTSC) subpopulations, co-hepato/pancreatic stem cells, involved in hepatic and pancreatic regeneration. FLCs and BTSCs express pluripotency genes, endodermal transcription factors, and stem cell surface, cytoplasmic and proliferation biomarkers.

The FLC-PDX model, FLC-TD-2010, is driven ex vivo to express pancreatic acinar traits, hypothesized responsible for this model's propensity for enzymatic degradation of cultures. A stable ex vivo model of FLC-TD-2010 was achieved using organoids in serum-free Kubota's Medium (KM) supplemented with 0.1% hyaluronans (KM/HA). Heparins (10 ng/ml) caused slow expansion of organoids with doubling times of ∼7–9 days. Spheroids, organoids depleted of mesenchymal cells, survived indefinitely in KM/HA in a state of growth arrest for more than 2 months. Expansion was restored with FLCs co-cultured with mesenchymal cell precursors in a ratio of 3:7, implicating paracrine signaling. Signals identified included FGFs, VEGFs, EGFs, Wnts, and others, produced by associated stellate and endothelial cell precursors.

Fifty-three, unique heparan sulfate (HS) oligosaccharides were synthesized, assessed for formation of high affinity complexes with paracrine signals, and each complex screened for biological activity(ies) on organoids. Ten distinct HS-oligosaccharides, all 10–12 mers or larger, and in specific paracrine signal complexes elicited particular biological responses. Of note, complexes of paracrine signals and 3-O sulfated HS-oligosaccharides elicited slowed growth, and with Wnt3a, elicited growth arrest of organoids for months. If future efforts are used to prepare HS-oligosaccharides resistant to breakdown in vivo, then [paracrine signal—HS-oligosaccharide] complexes are potential therapeutic agents for clinical treatments of FLCs, an exciting prospect for a deadly disease.

纤维板层癌 (FLC) 是一种发生在儿童到年轻人中的致命肿瘤，其遗传特征表明源自胆管树干细胞 (BTSC) 亚群，即肝/胰腺干细胞，参与肝脏和胰腺再生。FLC 和 BTSC 表达多能性基因、内胚层转录因子以及干细胞表面、细胞质和增殖生物标志物。

FLC-PDX 模型 FLC-TD-2010 被离体驱动以表达胰腺腺泡特征，推测是该模型对培养物进行酶促降解的倾向的原因。稳定的离体FLC-TD-2010 模型是使用补充有 0.1% 透明质酸 (KM/HA) 的无血清久保田培养基 (KM) 中的类器官实现的。肝素 (10 ng/ml) 导致类器官缓慢扩增，倍增时间约为 7-9 天。球状体，即耗尽间充质细胞的类器官，在 KM/HA 中无限期存活，并处于生长停滞状态超过 2 个月。FLC 与间充质细胞前体以 3:7 的比例共培养后，扩增恢复，这表明旁分泌信号传导。鉴定出的信号包括 FGF、VEGF、EGF、Wnt 等，由相关星状细胞和内皮细胞前体细胞产生。

合成了 53 种独特的硫酸乙酰肝素 (HS) 寡糖，评估了具有旁分泌信号的高亲和力复合物的形成，并筛选了每种复合物对类器官的生物活性。十种不同的 HS-寡糖，全部为 10-12 聚体或更大，并且在特定的旁分泌信号复合物中引发了特定的生物反应。值得注意的是，旁分泌信号和 3- O硫酸化 HS-寡糖的复合物引起生长减慢，并且与 Wnt3a 一起引起类器官生长停滞数月。如果未来努力制备在体内抗分解的HS-寡糖，那么[旁分泌信号-HS-寡糖]复合物将成为FLC临床治疗的潜在治疗剂，FLC是一种致命疾病的令人兴奋的前景。