Current Protocols in Stem Cell Biology最新文献_第10页

Human Adipose-Derived Stromal Cell Isolation Methods and Use in Osteogenic and Adipogenic In Vivo Applications 人脂肪来源的基质细胞分离方法及其在成骨和脂肪生成中的体内应用

Current Protocols in Stem Cell Biology Pub Date : 2018-02-13 DOI: 10.1002/cpsc.41

Elizabeth Brett, Ruth Tevlin, Adrian McArdle, Eun Young Seo, Charles K.F. Chan, Derrick C. Wan, Michael T. Longaker

{"title":"Human Adipose-Derived Stromal Cell Isolation Methods and Use in Osteogenic and Adipogenic In Vivo Applications","authors":"Elizabeth Brett, Ruth Tevlin, Adrian McArdle, Eun Young Seo, Charles K.F. Chan, Derrick C. Wan, Michael T. Longaker","doi":"10.1002/cpsc.41","DOIUrl":"10.1002/cpsc.41","url":null,"abstract":"Adipose tissue represents an abundant and easily accessible source of multipotent cells, which may serve as excellent building blocks for tissue engineering. This article presents a newly described protocol for isolating adipose-derived stromal cells (ASCs) from human lipoaspirate, compared to the standard protocol for harvesting ASCs established in 2001.Human ASC isolation is performed using two methods, and resultant cells are compared through cell yield, cell viability, cell proliferation and regenerative potential. The osteogenic and adipogenic potential of ASCs isolated using both protocols are assessed in vitro and gene expression analysis is performed. The focus of this series of protocols is the regenerative potential of both cell populations in vivo. As such, the two in vivo animal models described are fat graft retention (soft tissue reconstruction) and calvarial defect healing (bone regeneration). The techniques described comprise fat grafting with cell assisted lipotransfer, and calvarial defect creation healed with cell-seeded scaffolds. © 2017 by John Wiley & Sons, Inc.","PeriodicalId":53703,"journal":{"name":"Current Protocols in Stem Cell Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpsc.41","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35555094","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 6

Isolation of Ready-to-Use Adipose-Derived Stem Cell (ASC) Pellet for Clinical Applications and a Comparative Overview of Alternate Methods for ASC Isolation 用于临床应用的即食脂肪源性干细胞(ASC)颗粒的分离和ASC分离替代方法的比较综述

Current Protocols in Stem Cell Biology Pub Date : 2018-02-13 DOI: 10.1002/cpsc.29

Edoardo Raposio, Nicolò Bertozzi

{"title":"Isolation of Ready-to-Use Adipose-Derived Stem Cell (ASC) Pellet for Clinical Applications and a Comparative Overview of Alternate Methods for ASC Isolation","authors":"Edoardo Raposio, Nicolò Bertozzi","doi":"10.1002/cpsc.29","DOIUrl":"10.1002/cpsc.29","url":null,"abstract":"Current literature does not offer a standardized method to isolate adipose-derived stem cells (ASCs) for clinical applications and hence clinical studies using ASCs often show inconsistent results. Most of these studies borrow laboratory or benchside-derived protocols, which are complex, time consuming, and involve the use of chemical, animal-derived reagents. In this unit we describe a relatively simple and faster isolation protocol that allows collection of a ready-to-use ASC pellet for clinical application. All steps are performed in a closed circuit in order to guarantee maximum process sterility. Once the adipose tissue is harvested by means of a standard liposuction procedure, it undergoes a first centrifugation in order to remove the oil and serous fractions. Then ASCs are released by enzymatic digestion from the surrounding connective tissue scaffold. Finally a double series of washing and centrifugation allows one to obtain the ASC pellet alone. We usually graft this ASC pellet onto the skin edge and to the bottom of chronic skin ulcers as ASCs proved to be effective in promoting wound healing processes. Moreover, an increasing number of clinical studies are currently ongoing to test their potential in every medical field, from orthopedics to cardiology, oncology, autoimmune diseases, and tissue engineering. © 2017 by John Wiley & Sons, Inc.","PeriodicalId":53703,"journal":{"name":"Current Protocols in Stem Cell Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpsc.29","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35001879","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 41

Neural Stem Cell or Human Induced Pluripotent Stem Cell–Derived GABA-ergic Progenitor Cell Grafting in an Animal Model of Chronic Temporal Lobe Epilepsy 神经干细胞或人诱导多能干细胞衍生的gaba能祖细胞移植在慢性颞叶癫痫动物模型中的应用

Current Protocols in Stem Cell Biology Pub Date : 2018-02-13 DOI: 10.1002/cpsc.9

Dinesh Upadhya, Bharathi Hattiangady, Geetha A. Shetty, Gabriele Zanirati, Maheedhar Kodali, Ashok K. Shetty

{"title":"Neural Stem Cell or Human Induced Pluripotent Stem Cell–Derived GABA-ergic Progenitor Cell Grafting in an Animal Model of Chronic Temporal Lobe Epilepsy","authors":"Dinesh Upadhya, Bharathi Hattiangady, Geetha A. Shetty, Gabriele Zanirati, Maheedhar Kodali, Ashok K. Shetty","doi":"10.1002/cpsc.9","DOIUrl":"10.1002/cpsc.9","url":null,"abstract":"Grafting of neural stem cells (NSCs) or GABA-ergic progenitor cells (GPCs) into the hippocampus could offer an alternative therapy to hippocampal resection in patients with drug-resistant chronic epilepsy, which afflicts >30% of temporal lobe epilepsy (TLE) cases. Multipotent, self-renewing NSCs could be expanded from multiple regions of the developing and adult brain, human embryonic stem cells (hESCs), and human induced pluripotent stem cells (hiPSCs). On the other hand, GPCs could be generated from the medial and lateral ganglionic eminences of the embryonic brain and from hESCs and hiPSCs. To provide comprehensive methodologies involved in testing the efficacy of transplantation of NSCs and GPCs in a rat model of chronic TLE, NSCs derived from the rat medial ganglionic eminence (MGE) and MGE-like GPCs derived from hiPSCs are taken as examples in this unit. The topics comprise description of the required materials, reagents and equipment, methods for obtaining rat MGE-NSCs and hiPSC-derived MGE-like GPCs in culture, generation of chronically epileptic rats, intrahippocampal grafting procedure, post-grafting evaluation of the effects of grafts on spontaneous recurrent seizures and cognitive and mood impairments, analyses of the yield and the fate of graft-derived cells, and the effects of grafts on the host hippocampus. © 2016 by John Wiley & Sons, Inc.","PeriodicalId":53703,"journal":{"name":"Current Protocols in Stem Cell Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpsc.9","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34670819","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 23

Visualization and Modeling of the In Vivo Distribution of Mesenchymal Stem Cells 间充质干细胞体内分布的可视化和建模

Current Protocols in Stem Cell Biology Pub Date : 2018-02-13 DOI: 10.1002/cpsc.39

Haolu Wang, Camilla A. Thorling, Zhi Ping Xu, Darrell H. G. Crawford, Xiaowen Liang, Xin Liu, Michael S. Roberts

引用次数: 3

Mesenchymal Stem Cell Preparation and Transfection-free Ferumoxytol Labeling for MRI Cell Tracking 间充质干细胞制备和无转染阿魏木糖醇标记用于MRI细胞跟踪

Current Protocols in Stem Cell Biology Pub Date : 2018-02-13 DOI: 10.1002/cpsc.38

Li Liu, Chien Ho

引用次数: 11

CRISPR/Cas9-Based Safe-Harbor Gene Editing in Rhesus iPSCs 基于CRISPR/ cas9的安全港基因编辑在恒河猴iPSCs中的应用

Current Protocols in Stem Cell Biology Pub Date : 2018-02-13 DOI: 10.1002/cpsc.37

Ravi Chandra Yada, John W. Ostrominski, Ilker Tunc, So Gun Hong, Jizhong Zou, Cynthia E. Dunbar

引用次数: 6

Rapid Screening of the Endodermal Differentiation Potential of Human Pluripotent Stem Cells 人多能干细胞内胚层分化潜能的快速筛选

Current Protocols in Stem Cell Biology Pub Date : 2018-02-13 DOI: 10.1002/cpsc.36

Richard Siller, Gareth J. Sullivan

引用次数: 11

Generation of Oligodendrogenic Spinal Neural Progenitor Cells From Human Induced Pluripotent Stem Cells 从人诱导多能干细胞生成少突性脊神经祖细胞

Current Protocols in Stem Cell Biology Pub Date : 2017-08-14 DOI: 10.1002/cpsc.31

Mohamad Khazaei, Christopher S. Ahuja, Michael G. Fehlings

{"title":"Generation of Oligodendrogenic Spinal Neural Progenitor Cells From Human Induced Pluripotent Stem Cells","authors":"Mohamad Khazaei, Christopher S. Ahuja, Michael G. Fehlings","doi":"10.1002/cpsc.31","DOIUrl":"10.1002/cpsc.31","url":null,"abstract":"This unit describes protocols for the efficient generation of oligodendrogenic neural progenitor cells (o-NPCs) from human induced pluripotent stem cells (hiPSCs). Specifically, detailed methods are provided for the maintenance and differentiation of hiPSCs, human induced pluripotent stem cell-derived neural progenitor cells (hiPS-NPCs), and human induced pluripotent stem cell-oligodendrogenic neural progenitor cells (hiPSC-o-NPCs) with the final products being suitable for in vitro experimentation or in vivo transplantation. Throughout, cell exposure to growth factors and patterning morphogens has been optimized for both concentration and timing, based on the literature and empirical experience, resulting in a robust and highly efficient protocol. Using this derivation procedure, it is possible to obtain millions of oligodendrogenic-NPCs within 40 days of initial cell plating which is substantially shorter than other protocols for similar cell types. This protocol has also been optimized to use translationally relevant human iPSCs as the parent cell line. The resultant cells have been extensively characterized both in vitro and in vivo and express key markers of an oligodendrogenic lineage. © 2017 by John Wiley & Sons, Inc.","PeriodicalId":53703,"journal":{"name":"Current Protocols in Stem Cell Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpsc.31","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35268003","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 15

Generation of Xeno-Free, cGMP-Compliant Patient-Specific iPSCs from Skin Biopsy 从皮肤活检中生成无xeno、符合cgmp的患者特异性iPSCs

Current Protocols in Stem Cell Biology Pub Date : 2017-08-14 DOI: 10.1002/cpsc.30

Luke A. Wiley, Kristin R. Anfinson, Cathryn M. Cranston, Emily E. Kaalberg, Malia M. Collins, Robert F. Mullins, Edwin M. Stone, Budd A. Tucker

引用次数: 16

Functional Stem Cell Integration into Neural Networks Assessed by Organotypic Slice Cultures 器官型切片培养评估功能性干细胞与神经网络的整合

Current Protocols in Stem Cell Biology Pub Date : 2017-08-14 DOI: 10.1002/cpsc.34

David Forsberg, Charoensri Thonabulsombat, Johan Jäderstad, Linda Maria Jäderstad, Petri Olivius, Eric Herlenius

{"title":"Functional Stem Cell Integration into Neural Networks Assessed by Organotypic Slice Cultures","authors":"David Forsberg, Charoensri Thonabulsombat, Johan Jäderstad, Linda Maria Jäderstad, Petri Olivius, Eric Herlenius","doi":"10.1002/cpsc.34","DOIUrl":"10.1002/cpsc.34","url":null,"abstract":"Re-formation or preservation of functional, electrically active neural networks has been proffered as one of the goals of stem cell–mediated neural therapeutics. A primary issue for a cell therapy approach is the formation of functional contacts between the implanted cells and the host tissue. Therefore, it is of fundamental interest to establish protocols that allow us to delineate a detailed time course of grafted stem cell survival, migration, differentiation, integration, and functional interaction with the host. One option for in vitro studies is to examine the integration of exogenous stem cells into an existing active neural network in ex vivo organotypic cultures. Organotypic cultures leave the structural integrity essentially intact while still allowing the microenvironment to be carefully controlled. This allows detailed studies over time of cellular responses and cell-cell interactions, which are not readily performed in vivo. This unit describes procedures for using organotypic slice cultures as ex vivo model systems for studying neural stem cell and embryonic stem cell engraftment and communication with CNS host tissue. © 2017 by John Wiley & Sons, Inc.","PeriodicalId":53703,"journal":{"name":"Current Protocols in Stem Cell Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpsc.34","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35267931","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 7