Molecular perturbations restrict potential for liver repopulation of hepatocytes isolated from non-heart-beating donor rats.
Molecular perturbations restrict potential for liver repopulation of hepatocytes isolated from non-heart-beating donor rats.
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DOI：
10.1002/hep.24735
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发表时间：
2012-04
期刊：
影响因子：
13.5
通讯作者：
Gupta, Sanjeev
中科院分区：
文献类型：
--
作者：
Enami, Yuta;Joseph, Brigid;Bandi, Sriram;Lin, Juan;Gupta, Sanjeev
来自无心跳捐赠者的器官在细胞治疗中很有吸引力。了解再灌注/复氧后分子扰动的性质对于无心跳供体细胞将是非常重要的。我们研究了非心脏跳动供体大鼠的全球基因表达与Affyphin微阵列，肝组织的完整性，离体肝细胞的活力，移植细胞在二肽基肽酶IV缺陷大鼠的植入和增殖。在无心跳供体中，与有心跳供体相比，肝组织在死亡后4、16或34小时内形态完整，分别有1、95或372个基因的差异表达。这些差异表达的基因构成了氧化磷酸化、粘附连接、糖酵解/糖异生以及其他离散途径的本体途径中的突出分组。我们成功地从无心跳供体中分离出了活的肝细胞，特别是在死亡后4小时内，尽管肝细胞产量和存活率低于心跳供体的肝细胞，p<0.05。类似地，尽管来自无心跳供体的肝细胞在移植到受体动物中后移植并增殖，但这劣于来自心跳供体的肝细胞，p<0.05。与相应的肝组织相比，从无心跳供体中分离的肝细胞中的基因表达谱显示出更大的扰动，包括在粘着斑、肌动蛋白细胞骨架、细胞外基质-受体相互作用、多配体-受体相互作用中的途径的代表，以及胰岛素、钙、wnt、Jak-Stat或其他级联中的信号传导。结论：在非心脏跳动供体死亡后的长时间内，肝组织保持完整，但再灌注/复氧后广泛的分子扰动损害了这些供体分离的肝细胞的活力。对无心跳供体肝细胞分子变化的深入了解为提高供体细胞活力提供了机会，这将促进无心跳供体器官在细胞治疗或其他应用中的应用。
Organs from nonheart-beating donors are attractive for use in cell therapy. Understanding the nature of molecular perturbations following reperfusion/reoxygenation will be highly significant for nonheart-beating donor cells. We studied nonheart-beating donor rats for global gene expression with Affymetrix microarrays, hepatic tissue integrity, viability of isolated hepatocytes, and engraftment and proliferation of transplanted cells in dipeptidyl peptidase IV-deficient rats. In nonheart-beating donors, liver tissue was morphologically intact for >24 hours with differential expression of 1, 95, or 372 genes, 4, 16 or 34 hours after death, respectively, compared with heart-beating donors. These differentially-expressed genes constituted prominent groupings in ontological pathways of oxidative phosphorylation, adherence junctions, glycolysis/gluconeogenesis, as well as other discrete pathways. We successfully isolated viable hepatocytes from nonheart-beating donors, especially up to 4 hours after death, although the hepatocyte yield and viability were inferior to hepatocytes from heart-beating donors, p<0.05. Similarly, although hepatocytes from nonheart-beating donors engrafted and proliferated after transplantation in recipient animals, this was inferior to hepatocytes from heart-beating donors, p<0.05. Gene expression profiling in hepatocytes isolated from nonheart-beating donors showed far greater perturbations compared with corresponding liver tissue, including representation of pathways in focal adhesion, actin cytoskeleton, extracellular matrix-receptor interactions, multiple ligand-receptor interactions, and signaling in insulin, calcium, wnt, Jak-Stat, or other cascades. Conclusion: Liver tissue remained intact over prolonged periods after death in nonheart-beating donors but extensive molecular perturbations following reperfusion/reoxygenation impaired viability of isolated hepatocytes from these donors. Insights into molecular changes in hepatocytes from nonheart-beating donors offers opportunities for improving donor cell viability, which will advance utility of nonheart-beating donor organs for cell therapy or other applications.
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影响因子：
4.3
作者：
Calzado, Marco A.;De La Vega, Laureano;Schmitz, M. Lienhard
通讯作者：
Schmitz, M. Lienhard
影响因子：
50.3
作者：
Gregory MA;Phang TL;Neviani P;Alvarez-Calderon F;Eide CA;O'Hare T;Zaberezhnyy V;Williams RT;Druker BJ;Perrotti D;Degregori J
通讯作者：
Degregori J
影响因子：
13.5
作者：
Benten, D;Kumaran, V;Gupta, S
通讯作者：
Gupta, S
影响因子：
29.4
作者：
Enami, Yuta;Bandi, Sriram;Gupta, Sanjeev
通讯作者：
Gupta, Sanjeev
DOI：
10.1111/j.1432-1033.1996.0083t.x
发表时间：
1996-10-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Cai, Q;Storey, KB
通讯作者：
Storey, KB