Murine mammary stem/progenitor cell isolation: Different method matters?

doi:10.1186/s40064-016-1787-3

科研成果详情

题名	Murine mammary stem/progenitor cell isolation: Different method matters?
作者	Gao, Hui1,2; Dong, Qiaoxiang1,2,3; Chen, Yuanhong2,3; Zhang, Fuchuang2,3; Wu, Anqi2,3; Shi, Yuanshuo2,3; Bandyopadhyay, Abhik 2; Daniel, Benjamin J.4; Huang, Changjiang 3; Sun, Lu-Zhe 2
发表日期	2016-02-20
发表期刊	SPRINGERPLUS 影响因子和分区
语种	英语
原始文献类型	Article
关键词	Mammary stem/progenitor cell Enrichment Digestion method Surface marker Flow cytometric analysis
其他关键词	STEM-CELL ; EXPANSION ; GLAND
摘要	Murine mammary stem/progenitor cell isolation has been routinely used in many laboratories, yet direct comparison among different methods is lacking. In this study, we compared two frequently used digestion methods and three sets of frequently used surface markers for their efficiency in enriching mammary stem and progenitor cells in two commonly used mouse strains, C57BL/6J and FVB. Our findings revealed that the slow overnight digestion method using gentle collagenase/hyaluronidase could be easily adopted and yielded reliable and consistent results in different batches of animals. In contrast, the different fast digestion protocols, as described in published studies, yielded high percent of non-epithelial cells with very few basal epithelial cells liberated in our hands. The three sets of markers tested in our hands reveal rather equally efficiency in separating luminal and basal cells if same fluorochrome conjugations were used. However, the tendency of non-epithelial cell inclusion in the basal cell gate was highest in samples profiled by CD24/CD29 and lowest in samples profiled by CD49f/EpCAM, this is especially true in mammary cells isolated from C57BL/6J mice. This finding will have significant implication when sorted basal cells are used for subsequent gene expression analysis.
资助项目	National Natural Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [81373031]; National Institutes of HealthUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USA [R01 ES022057]; NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCESUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Institute of Environmental Health Sciences (NIEHS) [R01ES022057] Funding Source: NIH RePORTER
出版者	SPRINGER INTERNATIONAL PUBLISHING AG
出版地	CHAM
ISSN	2193-1801
卷号	5 期号:1 页码:140
DOI	10.1186/s40064-016-1787-3
页数	5
WOS类目	Multidisciplinary Sciences
WOS研究方向	Science & Technology - Other Topics
WOS记录号	WOS:000371414400005
收录类别	SCIE ; PUBMED ; SCOPUS
URL	查看原文
PubMed ID	26933638
PMC记录号	PMC4761358
SCOPUSEID	2-s2.0-84958756951
通讯作者地址	[Dong, Qiaoxiang]School of Laboratory Medicine and Life Science,Wenzhou Medical University,University Town,Wenzhou,325035,China
Scopus学科分类	Multidisciplinary
引用统计	被引频次：4[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	https://kms.wmu.edu.cn/handle/3ETUA0LF/4945
专题	检验医学院（生命科学学院、生物学实验教学中心）公共卫生学院_健康与环境生态研究所公共卫生学院_环境安全与健康风险评估研究院
通讯作者	Dong, Qiaoxiang
作者单位	1.School of Laboratory Medicine and Life Science,Wenzhou Medical University,University Town,Wenzhou,325035,China; 2.Department of Cellular and Structural Biology,University of Texas Health Science Center,San Antonio,78299,United States; 3.Institute of Environmental Safety and Human Health,Wenzhou Medical University,University Town,Wenzhou,325035,China; 4.Flow Cytometry Facility,University of Texas Health Science Center,San Antonio,78299,United States
第一作者单位	检验医学院（生命科学学院、生物学实验教学中心）
通讯作者单位	检验医学院（生命科学学院、生物学实验教学中心）
第一作者的第一单位	检验医学院（生命科学学院、生物学实验教学中心）
推荐引用方式 GB/T 7714	Gao, Hui,Dong, Qiaoxiang,Chen, Yuanhong,et al. Murine mammary stem/progenitor cell isolation: Different method matters?[J]. SPRINGERPLUS,2016,5(1):140.
APA	Gao, Hui., Dong, Qiaoxiang., Chen, Yuanhong., Zhang, Fuchuang., Wu, Anqi., ... & Sun, Lu-Zhe. (2016). Murine mammary stem/progenitor cell isolation: Different method matters?. SPRINGERPLUS, 5(1), 140.
MLA	Gao, Hui,et al."Murine mammary stem/progenitor cell isolation: Different method matters?".SPRINGERPLUS 5.1(2016):140.