Nevi Lorenzo, Carpino Guido, Costantini Daniele, Cardinale Vincenzo, Riccioni Olga, Di Matteo Sabina, Melandro Fabio, Berloco Pasquale Bartolomeo, Reid Lola, Gaudio Eugenio, Alvaro Domenico
Department of Medico-Surgical Sciences and Biotechnologies, Sapienza University of Rome, Rome, Italy.
Department of Movement, Human and Health Sciences, Division of Health Sciences, University of Rome "Foro Italico", Rome, Italy.
Stem Cell Res Ther. 2017 Mar 20;8(1):68. doi: 10.1186/s13287-017-0492-7.
Cell therapy of liver diseases with human biliary tree stem cells (hBTSCs) is biased by low engraftment efficiency. Coating the hBTSCs with hyaluronans (HAs), the primary constituents of all stem cell niches, could facilitate cell survival, proliferation, and, specifically, liver engraftment given that HAs are cleared selectively by the liver.
We developed a fast and easy method to coat hBTSCs with HA and assessed the effects of HA-coating on cell properties in vitro and in vivo.
The HA coating markedly improved the viability, colony formation, and population doubling of hBTSCs in primary cultures, and resulted in a higher expression of integrins that mediate cell attachment to matrix components. When HA-coated hBTSCs were transplanted via the spleen into the liver of immunocompromised mice, the engraftment efficiency increased to 11% with respect to 3% of uncoated cells. Notably, HA-coated hBTSC transplantation in mice resulted in a 10-fold increase of human albumin gene expression in the liver and in a 2-fold increase of human albumin serum levels with respect to uncoated cells. Studies in distant organs showed minimal ectopic cell distribution without differences between HA-coated and uncoated hBTSCs and, specifically, cell seeding in the kidney was excluded.
A ready and economical procedure of HA cell coating greatly enhanced the liver engraftment of transplanted hBTSCs and improved their differentiation toward mature hepatocytes. HA coating could improve outcomes of stem cell therapies of liver diseases and could be immediately translated into the clinic given that GMP-grade HAs are already available for clinical use.
人类胆管树干细胞（hBTSCs）用于肝脏疾病的细胞治疗受到低植入效率的限制。用透明质酸（HA）包被hBTSCs，而透明质酸是所有干细胞微环境的主要成分，鉴于肝脏可选择性清除透明质酸，这可能有助于细胞存活、增殖，尤其是肝脏植入。
我们开发了一种快速简便的方法用HA包被hBTSCs，并评估了HA包被对体外和体内细胞特性的影响。
HA包被显著提高了原代培养中hBTSCs的活力、集落形成和群体倍增，并导致介导细胞与基质成分附着的整合素表达更高。当将HA包被的hBTSCs经脾脏移植到免疫缺陷小鼠的肝脏中时，植入效率从未包被细胞的3%提高到了11%。值得注意的是，与未包被细胞相比，在小鼠中移植HA包被的hBTSC导致肝脏中人白蛋白基因表达增加了10倍，人白蛋白血清水平增加了2倍。对远处器官的研究表明异位细胞分布极少，HA包被和未包被的hBTSCs之间没有差异，特别是排除了细胞在肾脏中的定植。
一种现成且经济的HA细胞包被程序极大地增强了移植的hBTSCs在肝脏中的植入，并改善了它们向成熟肝细胞的分化。HA包被可以改善肝脏疾病干细胞治疗的效果，并且鉴于GMP级HA已可用于临床，可立即转化为临床应用。
