Tianjin Key Laboratory of Artificial Cell, Tianjin Institute of Hepatobiliary Disease, Artificial Cell Engineering Technology Research Center of Public Health Ministry, Third Central Hospital of Tianjin, Tianjin, China.
Department of Hepatobiliary Surgery, Third Central Hospital of Tianjin, Tianjin, China.
Cytotherapy. 2018 Jan;20(1):95-107. doi: 10.1016/j.jcyt.2017.08.008. Epub 2017 Sep 29.
Human induced pluripotent stem cells (hiPSCs) are becoming increasingly popular in research endeavors due to their potential for clinical application; however, such application is challenging due to limitations such as inferior function and low induction efficiency. In this study, we aimed to establish a three-dimensional (3D) culture condition to mimic the environment in which hepatogenesis occurs in vivo to enhance the differentiation of hiPSCs for large-scale culture and high throughput BAL application.
We used hydrogel to create hepatocyte-like cell (HLC) spheroids in a 3D culture condition and analyzed the cell-behavior and differentiation properties of hiPSCs in a synthetic nanofiber scaffold.
We found that treating cells with Y-27632 promoted the formation of spheroids, and the cells aggregated more rapidly in a 3D culture condition. The ALB secretion, urea production and glycogen synthesis by HLCs in 3D were significantly higher than those grown in a 2-dimensional culture condition. In addition, the metabolic activities of the CYP450 enzymes were also higher in cells differentiated in the 3D culture condition.
3D hydrogel culture condition can promote differentiation of hiPSCs into hepatocytes. The 3D culture approach could be applied to the differentiation of hiPSCs into hepatocytes for bioartificial liver.
由于人诱导多能干细胞（hiPSC）具有临床应用的潜力，因此在研究工作中越来越受欢迎；然而，由于功能低下和诱导效率低等限制，这种应用具有挑战性。在这项研究中，我们旨在建立一种三维（3D）培养条件，以模拟体内发生肝发生的环境，从而增强 hiPSC 的分化，以进行大规模培养和高通量 BAL 应用。
我们使用水凝胶在 3D 培养条件下创建肝细胞样细胞（HLC）球体，并在合成纳米纤维支架中分析 hiPSC 的细胞行为和分化特性。
我们发现用 Y-27632 处理细胞可促进球体的形成，并且细胞在 3D 培养条件下更迅速地聚集。与在 2D 培养条件下生长的细胞相比，3D 中 HLC 的 ALB 分泌、尿素产生和糖原合成明显更高。此外，在 3D 培养条件下分化的细胞中 CYP450 酶的代谢活性也更高。
3D 水凝胶培养条件可促进 hiPSC 分化为肝细胞。3D 培养方法可应用于 hiPSC 分化为肝细胞的生物人工肝脏。
