Cord Blood Expansion Inside a Bioengineered Liver

生物工程肝脏内的脐带血扩增

• 批准号: 8703780
• 负责人: Graca Duarte Almeida-Porada
• 金额: $ 11.32万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-19 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8703780
• 关键词: 3-Dimensional; Address; Adolescent; Adult; Behavior; Biliary; Biological Models; Biomedical Engineering; Bone Marrow; Cell Count; Cell Cycle; Cell Differentiation process; Cell Line; Cells; Confocal Microscopy; Cytoskeleton; Development; Disease; Elements; Endothelial Cells; Engraftment; Environment; Epithelial; Extracellular Matrix; Fetal Liver; Goals; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hepatic; Hepatic Tissue; Hereditary Disease; Human; Immune; In Vitro; Individual; Infection; Laboratories; Liver; Location; Lymphoid; Maintenance; Malignant - descriptor; Measures; Mesenchymal; Methylcellulose; Mus; Myelogenous; Non-Neoplastic Hematologic and Lymphocytic Disorder; Organoids; Patients; Phenotype; Physiological; Probability; Relative (related person); Research; Risk; Role; Serum; Sheep; Signal Transduction; Site; Source; Speed; Stem cells; Stromal Cells; Structure; System; Time; Tissues; Transplantation; Umbilical Cord Blood; Work; cancer genetics; cell type; clinically relevant; cytokine; fetal; functional outcomes; innovation; novel; novel strategies; progenitor; scaffold; self-renewal; stem

DESCRIPTION (provided by applicant): Cord blood (CB) is a clinically relevant source of hematopoietic stem/progenitor cells (HSPC) to treat cancer and genetic diseases. The advantages of using CB include its ready availability, the reduced probability of transmitting vira infections, and the lower risk of inducing graft vs. host disease in HLA-mismatched recipients. Still, CB's delayed speed of engraftment, and the relatively low number of hematopoietic stem cells (HSC) per unit, limit its broader use. Despite the advancements made in CB-HSPC expansion, challenges remain regarding the ability to obtain, from a single unit, sufficient numbers of both long-and short-term repopulating cells, for treatment of an adolescent or adult patient. We have previously shown that CB-HSPC can be expanded and differentiated towards both the myeloid and lymphoid lineages, using a feeder layer of adult human bone marrow-derived stromal cells. Using this system, we optimized the initial progenitor content and cytokine concentrations, and showed that expanded cells had the ability to engraft pre-immune fetal sheep. While the absolute number of long-term engrafting HSC increased in this culture system, still, the relative percentage of these most primitive stem cells decreasd with time. Recently, we have developed three- dimensional (3-D), liver extracellular matrix (ECM)-derived scaffolds and seeded them with fetal hepatoblasts and endothelial cells. These cells engrafted in their putative native locations within the liver ECM scaffolds, and subsequently displayed typical endothelial, hepatic, and biliary epithelial markers, thus creating a hepatic-lik tissue in vitro. It is well known that, during development, the fetal liver is the main site of HSC expansion and differentiation. Within the fetal liver, HSC actively cycle and these cells outcompete adult HSC upon transplantation. Thus, within the hepatic tissue, cellular niches exist that promote asymmetric or symmetric self-renewal divisions, leading to maintenance or expansion of primitive HSC. In addition, the initial divisional behavior of CB-HSPC is highly dependent upon the environment. For example, the stromal cell line AFT024 and fetal hepatoblasts, both of murine origin, have been shown, in 2-D cultures, to effectively preserve the self- renewal capacity of human and mouse HSC, respectively. Therefore, we hypothesize that a functional and efficient expansion of CB-HPSC can be achieved under physiological conditions provided by the bioengineered human hepatic constructs. Our ultimate goal is to develop a novel platform for the efficient expansion of CB- HSPC using bioengineered human liver tissue. To this end, we will: 1) Determine the ability of 3-D bioengineered huma liver tissue constructs to support ex-vivo expansion of CB-derived HSPC~ and 2) Examine and define the functional outcome arising from interactions that occur between CB-HSPC and individual cellular and matrix components of the niches of the bioengineered liver tissue. Upon completion, these studies will add to the understanding of how fetal liver niches support HSC expansion, and, more importantly, will allow the development of a novel strategy to functionally expand CB-HPSC.

描述（由申请人提供）：脐带血（CB）是临床相关的造血干细胞/祖细胞（HSPC）来源，用于治疗癌症和遗传疾病。使用CB的优点包括其现成性、传播病毒感染的可能性降低以及在HLA不匹配的受者中诱发移植物抗宿主病的风险较低。 尽管如此，CB 的植入速度较慢，且每单位造血干细胞 (HSC) 数量相对较低，限制了其更广泛的使用。 尽管在 CB-HSPC 扩增方面取得了进展，但从单个单元获得足够数量的长期和短期再生细胞用于治疗青少年或成年患者的能力仍然存在挑战。 我们之前已经证明，使用成人骨髓源性基质细胞的饲养层，CB-HSPC 可以向骨髓和淋巴谱系扩展和分化。使用该系统，我们优化了初始祖细胞含量和细胞因子浓度，并表明扩增的细胞具有植入免疫前胎羊的能力。 虽然在该培养系统中长期移植的 HSC 的绝对数量有所增加，但这些最原始干细胞的相对百分比却随着时间的推移而减少。 最近，我们开发了三维（3-D）肝脏细胞外基质（ECM）衍生的支架，并在其中接种了胎儿肝母细胞和内皮细胞。这些细胞被移植到肝脏 ECM 支架内假定的天然位置，随后 显示出典型的内皮、肝和胆管上皮标志物，从而在体外形成肝样组织。众所周知，在发育过程中，胎儿肝脏是HSC的主要场所。 扩张和分化。 在胎儿肝脏内，HSC 活跃地循环，并且这些细胞在移植后会击败成人 HSC。因此，在肝组织内，存在促进不对称或对称自我更新分裂的细胞生态位，从而导致原始HSC的维持或扩张。此外，CB-HSPC 的初始分裂行为高度依赖于环境。例如，小鼠来源的基质细胞系 AFT024 和胎儿成肝细胞在二维培养物中已被证明能够分别有效地保留人和小鼠 HSC 的自我更新能力。 因此，我们假设在生物工程人肝构建体提供的生理条件下可以实现 CB-HPSC 的功能性和有效扩展。我们的最终目标是开发一个新的平台，利用生物工程人体肝脏组织有效扩展 CB-HSPC。 为此，我们将：1) 确定 3-D 生物工程人肝组织构建体支持 CB 衍生的 HSPC~ 离体扩增的能力；2) 检查并定义 CB- 之间发生的相互作用所产生的功能结果HSPC 以及生物工程肝组织微环境的单个细胞和基质成分。完成后，这些研究将加深对胎儿肝脏生态位如何支持 HSC 扩增的理解，更重要的是，将允许开发一种新的策略来功能性地扩展 CB-HPSC。