Department of Surgery, Geneva University Hospitals and Medical Faculty, 1211 Geneva, Switzerland.
Institute of Chemical Sciences and Engineering, Ecole Polytechnique Fédérale de Lausanne, 1015 Lausanne, Switzerland.
J Immunol Res. 2018 Jan 11;2018:1078547. doi: 10.1155/2018/1078547. eCollection 2018.
Porcine hepatocytes transplanted during acute liver failure might support metabolic functions until the diseased liver recovers its function. Here, we isolated high numbers of viable pig hepatocytes and evaluated hepatocyte functionality after encapsulation. We further investigated whether coculture and coencapsulation of hepatocytes with human multipotent mesenchymal stromal cells (MSC) are beneficial on hepatocyte function. Livers from 10 kg pigs ( = 9) were harvested, and hepatocytes were isolated from liver suspensions for microencapsulation using alginate and poly(ethylene-glycol)- (PEG-) grafted alginate hydrogels, either alone or in combination with MSC. Viability, albumin secretion, and diazepam catabolism of hepatocytes were measured for one week. 9.2 ± 3.6 × 10 hepatocytes with 95.2 ± 3.1% viability were obtained after isolation. At day 3, free hepatocytes displayed 99% viability, whereas microencapsulation in alginate and PEG-grafted alginate decreased viability to 62% and 48%, respectively. Albumin secretion and diazepam catabolism occurred in free and microencapsulated hepatocytes. Coencapsulation of hepatocytes with MSC significantly improved viability and albumin secretion at days 4 and 8 ( < 0.05). Coculture with MSC significantly increased and prolonged albumin secretion. In conclusion, we established a protocol for isolation and microencapsulation of high numbers of viable pig hepatocytes and demonstrated that the presence of MSC is beneficial for the viability and function of porcine hepatocytes.
在急性肝功能衰竭期间移植的猪肝细胞可能会支持代谢功能,直到病变的肝脏恢复其功能。在这里,我们分离出大量有活力的猪肝细胞,并评估了包封后的肝细胞功能。我们进一步研究了猪肝细胞与人类多能间充质基质细胞 (MSC) 共培养和共包封是否对肝细胞功能有益。从 10kg 猪的肝脏 ( = 9) 中收获肝脏,并从肝混悬液中分离肝细胞,用于使用藻酸盐和聚(乙二醇)接枝藻酸盐水凝胶进行微囊化,单独或与 MSC 一起使用。测量了肝细胞的活力、白蛋白分泌和地西泮代谢。分离后获得了 9.2 ± 3.6 × 10 个具有 95.2 ± 3.1%活力的肝细胞。第 3 天,游离肝细胞的活力为 99%,而藻酸盐和聚乙二醇接枝藻酸盐微囊化分别将活力降低至 62%和 48%。游离和微囊化的肝细胞均能进行白蛋白分泌和地西泮代谢。将 MSC 与肝细胞共包封可显著提高第 4 天和第 8 天的活力和白蛋白分泌( < 0.05)。与 MSC 共培养可显著增加并延长白蛋白分泌。总之,我们建立了一个从猪肝细胞中分离和微囊化大量有活力的猪肝细胞的方案,并证明 MSC 的存在有利于猪肝细胞的活力和功能。