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比较小鼠原代肝细胞和胚胎肝祖细胞在体外的增殖和分化潜能。

Comparison of proliferation and differentiation potential between mouse primary hepatocytes and embryonic hepatic progenitor cells in vitro.

机构信息

Department of Pediatric Surgery, Stem Cell Biology and Therapy Laboratory, Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing Stem Cell Therapy Engineering Technical Center, The Children's Hospital of Chongqing Medical University, Chongqing 400014, P.R. China.

出版信息

Int J Mol Med. 2013 Aug;32(2):476-84. doi: 10.3892/ijmm.2013.1413. Epub 2013 Jun 11.

Abstract

Cell therapy may be a novel and effective treatment strategy for liver diseases, replacing liver transplantation. The potential of two alternative cell types (hepatic progenitor/stem cells and mature hepatocytes) has not yet been fully assessed; the issues of low amplification efficiency and recovery function remain to be resolved. In this study, we investigated the proliferation, differentiation and function of primary mouse mature hepatocytes and embryonic hepatic progenitor cells. Primary cells were obtained from the livers of mouse embryos at 14.5 days post coitus [hepatic progenitor 14.5d (HP14.5d) cells], as well as from the livers of 3-month-old mice [liver cells 3m (LC3m)]. Using trypan blue staining and crystal violet staining to detect cell viability, we found that compared with the limited growth capability of primary LC3m cells, primary HP14.5d cells exhibited an active cell proliferation; however, proliferative ability of passaged HP14.5d cells significantly decreased. After the HP14.5d cells were treated in hepatic induction medium, the expression of progenitor cell markers decreased and that of mature hepatic markers increased, to levels similar to those of LC3m cells. On day 12 of induction, the HP14.5d cells showed comparable indocyanine green (ICG) uptake and glycogen storage to that of the LC3m cells. Therefore, our study demonstrates that primary hepatic progenitor cells have a stronger proliferation capacity and differentiation potential, supporting their clinical application in liver cell transplantation.

摘要

细胞治疗可能是一种新颖且有效的肝脏疾病治疗策略,可以替代肝移植。两种替代细胞类型(肝祖细胞/干细胞和成熟肝细胞)的潜力尚未得到充分评估;扩增效率低和恢复功能的问题仍有待解决。在这项研究中,我们研究了原代小鼠成熟肝细胞和胚胎肝祖细胞的增殖、分化和功能。原代细胞从合胞体后 14.5 天的小鼠胚胎肝脏中获得(肝祖细胞 14.5d [HP14.5d] 细胞),以及从 3 个月大的小鼠肝脏中获得(LC3m 细胞)。通过台盼蓝染色和结晶紫染色检测细胞活力,我们发现与原代 LC3m 细胞有限的生长能力相比,原代 HP14.5d 细胞表现出活跃的细胞增殖;然而,传代 HP14.5d 细胞的增殖能力显著降低。在 HP14.5d 细胞用肝诱导培养基处理后,祖细胞标志物的表达减少,成熟肝标志物的表达增加,与 LC3m 细胞相似。在诱导的第 12 天,HP14.5d 细胞表现出与 LC3m 细胞相当的吲哚菁绿(ICG)摄取和糖原储存能力。因此,我们的研究表明,原代肝祖细胞具有更强的增殖能力和分化潜力,支持它们在肝细胞移植中的临床应用。

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