优化生长因子诱导人多能干细胞高效生成肝细胞样细胞。

IF 0.3 Q4 TRANSPLANTATION
International Journal of Organ Transplantation Medicine Pub Date : 2018-01-01 Epub Date: 2018-05-01
Z Jafarpour, M Soleimani, S Hosseinkhani, M H M H, P Yaghmaei, N Mobarra, B Geramizadeh
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引用次数: 0

摘要

背景:生成具有完整肝功能的肝细胞仍然是一个挑战,需要开发更多的功能肝细胞。目的:比较人诱导多能干细胞(human induced pluripotent stem cells, hiPSCs)向肝细胞样细胞(hepatocyte-like cells, HLCs)分化的各种分化因子和分化方案,提出一种较好的分化方案。方法:hiPSCs内胚层分化3 d后,用5种肝细胞分化培养基(P)孵育14 d - p1:第一周用肝细胞生长因子和成纤维细胞生长因子-4 (FGF-4),第二周用癌他汀- m和地塞米松;P2:与P1相似,但第一周和第二周均使用FGF4;P3:与P1相似,但未使用FGF-4;P4:与P1相似,但未使用FGF-4和地塞米松;P5:与P1相似,但不使用FGF-4和oncostatin-M。17 d后,采用qRT-PCR、免疫荧光和ELISA进行鉴定。结果:肝细胞标志物(白蛋白、细胞角蛋白-18、酪氨酸转氨酶、肝细胞核因子-4α、细胞色素- p450 7A1) mRNA表达水平显著升高(p)。结论:P3在所有方案中效果最好,我们推荐P3作为区分功能性hlc和hiPSCs的有效方案,可以改善细胞治疗。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Efficient Production of Hepatocyte-like Cells from Human-induced Pluripotent Stem Cells by Optimizing Growth Factors.

Efficient Production of Hepatocyte-like Cells from Human-induced Pluripotent Stem Cells by Optimizing Growth Factors.

Efficient Production of Hepatocyte-like Cells from Human-induced Pluripotent Stem Cells by Optimizing Growth Factors.

Efficient Production of Hepatocyte-like Cells from Human-induced Pluripotent Stem Cells by Optimizing Growth Factors.

Background: Generating hepatocytes with complete liver functions is still a challenge and developing more functional hepatocytes is needed.

Objective: To compare various differentiation factors and protocols and introducing a preferable protocol to differentiate human-induced pluripotent stem cells (hiPSCs) into hepatocyte-like cells (HLCs).

Methods: After 3 days of the endoderm differentiation of hiPSCs, the cells were incubated with 5 hepatocyte differentiation culture media, protocols (P), for 14 days-P1: hepatocyte growth factor and fibroblast growth factor-4 (FGF-4) for the first week and oncostatin-M and dexamethasone for the second week; P2: similar to P1 but FGF4 was used in both the first and second weeks; P3: similar to P1 but FGF-4 was not used; P4: similar to P1 but FGF-4 and dexamethasone were not used; and P5: similar to P1 but FGF-4 and oncostatin-M were not used. After 17 days, characterization was done by qRT-PCR, immunofluorescence and ELISA.

Results: The mRNA expression levels of hepatocyte markers (albumin, cytokeratin-18, tyrosine aminotransferase, hepatocyte nuclear factor-4α, cytochrome-P450 7A1) increased significantly (p<0.05) in the differentiated cells by 5 different protocols. Furthermore, significant protein expression and secretion of albumin were detected in the differentiated cells by 5 different protocols. In P3, the differentiated cells had the highest exhibit of hepatocyte characteristics and in P4 they had the lowest. Moreover, in P1 and P2 similar results were observed.

Conclusion: Since P3 gave us the best results among all protocols, we recommend it as an efficient protocol to differentiate the functional HLCs from hiPSCs, which can improve cell therapies.

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来源期刊
CiteScore
1.60
自引率
0.00%
发文量
0
审稿时长
12 weeks
期刊介绍: The International Journal of Organ Transplantation Medicine (IJOTM) is a quarterly peer-reviewed English-language journal that publishes high-quality basic sciences and clinical research on transplantation. The scope of the journal includes organ and tissue donation, procurement and preservation; surgical techniques, innovations, and novelties in all aspects of transplantation; genomics and immunobiology; immunosuppressive drugs and pharmacology relevant to transplantation; graft survival and prevention of graft dysfunction and failure; clinical trials and population analyses in the field of transplantation; transplant complications; cell and tissue transplantation; infection; post-transplant malignancies; sociological and ethical issues and xenotransplantation.
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