Adult Stem Cell Enrichment via Biomechanical Characterization

描述（由申请人提供）：脂肪来源的成体基质/干（ADAS）细胞为生物医学工程领域带来了令人兴奋的机会。ADAS细胞作为组织再生程序的细胞来源是有吸引力的，因为它们容易获得和有限的供体部位发病率。过去的研究表明，这些细胞可以使用培养基补充剂分化成各种细胞类型，有效地引导它们朝向感兴趣的特定谱系：软骨细胞，成骨细胞和脂肪细胞。本实验室目前的研究表明，完全分化的细胞和未分化的ADAS细胞具有其细胞类型（成骨细胞、软骨细胞、脂肪细胞）的生物力学特性。该项目指导阶段的目标是确定干细胞分化过程中细胞力学如何变化。我们假设细胞生物力学特性反映了干细胞分化的阶段，并为细胞分化为特定表型的潜力提供了一个标记。为了测试这一点，我们将使用原子力显微镜确定单个ADAS细胞在分化过程中的弹性和粘弹性。相比 原代细胞类型将有助于确定ADAS细胞是否在细胞分化方面成功分化， 生物力学此外，量化细胞的局部力学将有助于理解这些组织中的力学转导。指导阶段的研究将显着提高我们对干细胞分化过程中发生的细胞水平变化的理解。直接向建议的研究为独立阶段的奖励。该阶段的目标是通过其生物力学特性分离脂肪抽吸物中的细胞，以产生将改善工程组织性能的谱系特异性细胞来源。我们假设，含有高百分比的组织特异性干细胞的工程软骨将具有更好的机械和生物化学特性比接种更异质的收获群体的结构。为了测试这一点，我们将从吸脂废物组织中分选细胞，通过它们的生物力学特性，随后评估它们的多能性。组织工程软骨将使用正常和分选的细胞群生长，以确定干细胞富集如何影响组织形成的质量。整个拟议的项目将在机械传导，干细胞生物学和组织工程领域取得重大进展。

DESCRIPTION (provided by applicant): Adipose-derived adult stromal/stem (ADAS) cells pose exciting opportunities for the field of biomedical engineering. ADAS cells are attractive as a cell source for tissue regeneration procedures because of their ready availability and limited donor-site morbidity. Past research has shown that these cells can be differentiated into a variety of cell types using media supplements, effectively guiding them towards specific lineages of interest: chondrocytic, osteoblastic, and adipocytic. Current studies in our laboratory indicate that fully differentiated cells' and undifferentiated ADAS cells posses biomechanical properties characteristic of their cell type (osteoblast, chondrocyte, adipocyte). The goal of the Mentored phase of the project is to determine how cellular mechanics change during stem cell differentiation. We hypothesize that cellular biomechanical properties reflect the stage of stem cell differentiation and provide a marker for the cell's potential to differentiate into specific phenotypes. To test this, we will determine the elastic and viscoelastic properties of single, ADAS cells during differentiation using atomic force microscopy. Comparison with primary cell types will help determine whether ADAS cells successfully differentiate in terms of cellular biomechanics. Furthermore, quantifying the local mechanics of cells will help in understanding mechanotransduction in these tissues. The Mentored phase studies will significantly improve our understanding of the cellular-level changes that occur during the stem cell differentiation process and lead. Directly to the proposed research for the Independent phase of the award. The goal of this phase is to separate cells within lipoaspirate by their biomechanical properties to produce lineage-specific cell sources that will improve the performance of engineered tissues. We hypothesize that engineered cartilage containing a high percentage of tissue-specific stem cells will possess better mechanical and biochemical properties than constructs seeded with a more heterogeneous harvest population. To test this, we will sort cells from liposuction waste tissue by their biomechanical properties and subsequently evaluate their multipotent character. Tissue engineered cartilage will be grown using normal and sorted cell populations to determine how stem cell enrichment affects the quality of tissue formation. The entire proposed project will produce significant advances in the areas of mechanotransduction, stem cell biology, and tissue engineering.