Control of Stem Cell Fate in Heart Coculture

描述（申请人提供）：心血管疾病是美国最使人衰弱的疾病之一。目前，导致发育异常或患病心脏修复的机制尚不清楚。心脏细胞替代在移植研究中取得了令人鼓舞的结果。然而，目前尚不清楚哪种细胞类型或微环境将是最有效的细胞替换。本提案将检查人类胚胎干细胞（hES）的能力，NIH注册号：WA01和胚胎生殖细胞（hEG）用于修复或再生发育异常或受损的心脏模型。在目的1中，器官共培养将用于为胚胎7-12天小鼠心脏的心肌细胞分化创造类似于体内的环境。Aim 2将检测人类胚胎干细胞或EG细胞在Gja1基因突变的小鼠（具有先天性心脏异常）中重新填充心脏干细胞生态位的能力，以及修复被白喉毒素损伤的心脏组织的能力，以确定心脏干细胞生态位在指导hEG或hES分化中的作用。在Aim 3中，将心脏神经嵴细胞与hES或hEG细胞共培养，以确定这些细胞是否也能指导心肌细胞分化。

DESCRIPTION (provided by applicant): Cardiovascular disease is one of the most debilitating diseases in America. Currently, mechanisms leading to repair of developmentally abnormal or diseased hearts is unknown. Encouraging results in transplantation studies have been shown with cardiac cell replacement. However, it is not known which cell types or microenvironment will be the most efficient for cell replacement. This proposal will examine the ability of human embryonic stem (hES), NIH registry no. WA01, and embryonic germ cells (hEG) for repair or repopulation of developmentally abnormal or damaged heart models. In Aim 1, organ coculture will be used to create an environment similar to in vivo for differentiation of cardiomyocytes in embryonic day 7-12 mouse hearts. Aim 2 will examine the ability of human ES or EG cells to repopulate the cardiac stem cell niche in mice with a mutation in the Gja1 gene, which have congenital cardiac abnormalities, and also to repair heart tissue damaged by diptheria toxin to determine the role of the cardiac stem cell niche in directing hEG or hES differentiation. In Aim 3, coculture of cardiac neural crest cells with hES or hEG cells will be used to determine if these cells may also be able to direct cardiomyocyte differentiaton.