Ex Vivo Development of Transfected Oral Mucosal Grafts

转染口腔粘膜移植物的离体发育

• 批准号: 7578240
• 负责人: STEPHEN Elliott FEINBERG
• 金额: $ 34.78万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-01 至 2010-08-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7578240
• 关键词: Adult; Animal Model; Biological Assay; Canis familiaris; Cell Proliferation; Cell Size; Cells; Clinical Trials; Data; Defect; Development; Devices; FDA approved; Goals; Guidelines; Human; Immunocompromised Host; Lead; Modeling; Nuclear; Oral; Oral cavity; Oral mucous membrane structure; Pathway interactions; Patient Care; Population; Replacement Therapy; Research Personnel; Research Project Grants; SCID Mice; Safety; Sorting - Cell Movement; Specific qualifier value; Stem Cell Research; Stem cells; Suspension substance; Suspensions; Technology; Tissue Engineering; base; bench to bedside; gene therapy; improved; keratinocyte; oral mucosal graft; progenitor; programs; reconstruction; stem cell population; success; transcription factor; translational study

DESCRIPTION: Our long-term goal is to produce a "smart" transduced oral mucosal graft that will be used for reconstruction of major oral defects. Currently, an unfractionated unsorted human adult single keratinocyte suspension is used to manufacture a "naive" non-transduced Ex Vivo Produced Oral Mucosa Equivalent (EVPOME) that is suitable to perform FDA-approved human clinical trials. Our success with fractionated unsorted cultured adult human oral mucosal cells provides compelling empiric evidence that a subpopulation exists within our cell population that represents the "stem" cell compartment. In order for our technology/gene therapy to become a clinically viable option we need to isolate a progenitor/stem cell population. Our hypothesis states: "A subpopulation of fractionated sorted oral mucosa keratinocytes enriched for progenitor/stem cells can be: 1) isolated and 2) manipulated to alter their replication and differentiation, to enhance their ability to fabricate a superior EVPOME graft for intraoral grafting and for use as a device for gene therapy. Specific Aims: 1. Isolate and characterize an "enriched population" of oral mucosa progenitor/stem cells from fractionated sorted cultured primary human oral keratinocytes. 2. Demonstrate the safety and fate of a transduced enriched population of oral mucosa progenitor/stem cells in a "smart" EVPOME by grafting into both SCID mouse and canine models. 3. Alter expression of nuclear transcription factor PPARy to enhance replication and decrease differentiation of an enriched population of oral mucosa progenitor/stem cells. This research project is a logical and necessary iteration of the "bench to bedside" paradigm as it applies to the EVPOME. The results to establish expanded cultures of an enriched population of oral mucosa progenitor/stem cells, under chemically defined conditions and FDA guidelines, can lead to greater advances in cell replacement therapy. These types of translational studies are needed to generate new cell based tissue-engineered devices to improve patient care.

描述：我们的长期目标是生产一种“智能”的转导口腔粘膜移植物，将用于重建主要的口腔缺损。目前，使用未分级的未分选的成人单个角质形成细胞悬浮液来制造适合于进行FDA批准的人类临床试验的“幼稚”非转导的离体产生的口腔粘膜等同物（EVPOME）。我们的成功与分级未分选培养的成人口腔粘膜细胞提供了令人信服的经验证据表明，一个亚群存在于我们的细胞群，代表“干”细胞室。为了使我们的技术/基因疗法成为临床上可行的选择，我们需要分离祖细胞/干细胞群体。 我们的假设是：富集祖细胞/干细胞的分级分选的口腔粘膜角质形成细胞的亚群可以：1）分离和2）操作以改变它们的复制和分化，以增强它们制造用于口内移植和用作基因治疗装置的上级EVPOME移植物的能力。具体目标： 1.从分级分选的培养的原代人口腔角质形成细胞中分离并表征口腔粘膜祖细胞/干细胞的“富集群体”。 2.通过移植到SCID小鼠和犬模型中，证明“智能”EVPOME中口腔粘膜祖细胞/干细胞的转导富集群体的安全性和命运。 3.改变核转录因子PPARy的表达以增强口腔粘膜祖细胞/干细胞富集群体的复制并降低其分化。 该研究项目是“从工作台到床边”范式的逻辑和必要迭代，因为它适用于EVPOME。在化学定义的条件和FDA指南下建立口腔粘膜祖细胞/干细胞富集群体的扩增培养物的结果可以导致细胞替代疗法的更大进步。需要这些类型的转化研究来产生新的基于细胞的组织工程装置，以改善患者护理。

项目成果

Non-linear stress-strain measurements of ex vivo produced oral mucosal equivalent (EVPOME) compared to normal oral mucosal and skin tissue.

与正常口腔粘膜和皮肤组织相比，对离体产生的口腔粘膜当量（EVPOME）进行非线性应力应变测量。

• DOI: 10.1109/iembs.2011.6090075
• 发表时间: 2011
• 期刊: Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Annual International Conference
• 作者: Winterroth,Frank;Hollister,ScottJ;Feinberg,StephenE;Kuo,Shiuhyang;Fowlkes,JBrian;Ganguly,Arindam;Hollman,KyleW
• 通讯作者: Hollman,KyleW

Ex vivo produced human conjunctiva and oral mucosa equivalents grown in a serum-free culture system.

离体产生在无血清培养系统中生长的人类结膜和口腔粘膜等同物。

• DOI: 10.1016/j.joms.2004.02.010
• 发表时间: 2004
• 期刊: Journal of oral and maxillofacial surgery : official journal of the American Association of Oral and Maxillofacial Surgeons
• 作者: Yoshizawa,Michiko;Feinberg,StephenE;Marcelo,CynthiaL;Elner,VictorM
• 通讯作者: Elner,VictorM

Isolation of small-sized human epidermal progenitor/stem cells by Gravity Assisted Cell Sorting (GACS).

• DOI: 10.1016/j.jdermsci.2009.09.003
• 发表时间: 2009-12
• 期刊: Journal of dermatological science
• 影响因子: 4.6
• 作者: Y. Fujimori;K. Izumi;S. Feinberg;C. Marcelo

Characterization of a unique technique for culturing primary adult human epithelial progenitor/"stem cells".

• DOI: 10.1186/1471-5945-12-8
• 发表时间: 2012-06-24
• 期刊: BMC dermatology
• 作者: Marcelo CL;Peramo A;Ambati A;Feinberg SE
• 通讯作者: Feinberg SE

Constitutive release of cytokines by human oral keratinocytes in an organotypic culture.

器官型培养物中人口腔角质形成细胞持续释放细胞因子。

• DOI: 10.1016/j.joms.2009.02.003
• 发表时间: 2009
• 期刊: Journal of oral and maxillofacial surgery : official journal of the American Association of Oral and Maxillofacial Surgeons
• 作者: Xu,Qin;Izumi,Kenji;Tobita,Takayoshi;Nakanishi,Yoshitaka;Feinberg,StephenE
• 通讯作者: Feinberg,StephenE