純小分子雞尾酒療法促進人類多能幹細胞向肝細胞的分化

論文標題：Highly efficient and expedited hepatic differentiation from human pluripotent stem cells by pure small-molecule cocktails

數字識別碼：10.1186/s13287-018-0794-4

原文作者：Cong Du, Yuan Feng, Dongbo Qiu, Yan Xu, Mao Pang, Nan Cai, Andy Peng Xiang and Qi Zhang

人類誘導多能幹細胞的出現為產生充足的各異的肝細胞帶來了巨大希望。儘管之前已有工作通過基於病毒的轉錄因子和/或在分化過程中添加生長因子等技術在體外成功地從人多能幹細胞產生了肝細胞，但病毒轉導的安全性問題和細胞因子的高成本阻礙了其下游的應用。最近，由於具有優越的穩定性、安全性、細胞滲透性和成本效益，小分子的使用成為誘導細胞命運轉變的一種有力工具。

在發表於Stem Cell Research & Therapy的Highly efficient and expedited hepatic differentiation from human pluripotent stem cells by pure small-molecule cocktails一文中，來自中山大學附屬第三醫院的Qi Zhang及其研究團隊基於純小分子雞尾酒療法，開發了一種新型高效的人類多能幹細胞的肝細胞分化方案。這種方法依靠三步策略在短短的13天內，逐步誘導肝細胞的分化，包括定型內胚層分化、肝臟特化和肝細胞成熟。

圖1. 誘導hPSC逐步分化為肝細胞的三階段。

Cong Du et al.

分化的肝細胞樣細胞在形態學上類似於基於生長因子法產生的肝細胞和原代肝細胞。這些細胞不僅在轉錄和蛋白水平上表達特異性肝臟標誌物，還具有主要的肝功能，例如白蛋白產生、糖原儲存、細胞色素P450的活性和吲哚菁綠的吸收和釋放。

通過這種新穎的純小分子雞尾酒方法，可以實現人多能幹細胞高效快速的肝細胞分化，這為體外研究人類肝臟發育的分子機制提供了一個經濟有效的平台，在未來臨床應用方面有巨大的潛力。

摘要：

Background

The advent of human-induced pluripotent stem cells holds great promise for producing ample individualized hepatocytes. Although previous efforts have succeeded in generating hepatocytes from human pluripotent stem cells in vitro by viral-based expression of transcription factors and/or addition of growth factors during the differentiation process, the safety issue of viral transduction and high cost of cytokines would hinder the downstream applications. Recently, the use of small molecules has emerged as a powerful tool to induce cell fate transition for their superior stability, safety, cell permeability, and cost-effectiveness.

Methods

In the present study, we established a novel efficient hepatocyte differentiation strategy of human pluripotent stem cells with pure small-molecule cocktails. This method induced hepatocyte differentiation in a stepwise manner, including definitive endoderm differentiation, hepatic specification, and hepatocyte maturation within only 13 days.

Results

The differentiated hepatic-like cells were morphologically similar to hepatocytes derived from growth factor-based methods and primary hepatocytes. These cells not only expressed specific hepatic markers at the transcriptional and protein levels, but also possessed main liver functions such as albumin production, glycogen storage, cytochrome P450 activity, and indocyanine green uptake and release.

Conclusions

Highly efficient and expedited hepatic differentiation from human pluripotent stem cells could be achieved by our present novel, pure, small-molecule cocktails strategy, which provides a cost-effective platform for in vitro studies of the molecular mechanisms of human liver development and holds significant potential for future clinical applications.

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期刊介紹:

Stem Cell Research & Therapy(https://stemcellres.biomedcentral.com/ ,4.963 -2-year Impact Factor, 5.382 -5-year Impact Factor) is the major forum for translational research into stem cell therapies. An international peer-reviewed journal, it publishes high-quality open access research articles with a special emphasis on basic, translational and clinical research into stem cell therapeutics and regenerative therapies, including animal models and clinical trials. The journal also provides reviews, viewpoints, commentaries, reports and methods.

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