北京大学医学部机构知识库
Advanced  
IR@PKUHSC  > 北京大学第一临床医学院  > 期刊论文
学科主题: 临床医学
题名:
Biostorage and Quality Control for Human Peripheral Blood Leukocytes
作者: Zhou, Jiayi1; Wu, Jing1; Zhao, Xiaoyan1,2; Shen, Weihong1; Liu, Xiaoyan1; Xu, Congjian1,2; Jin, Hongyan1,2,3
刊名: BIOPRESERVATION AND BIOBANKING
发表日期: 2015-02-01
DOI: 10.1089/bio.2014.0094
卷: 13, 期:1, 页:13-19
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Cell Biology ; Chemistry, Applied ; Medical Laboratory Technology
研究领域[WOS]: Cell Biology ; Chemistry ; Medical Laboratory Technology
关键词[WOS]: DNA STORAGE ; TEMPERATURE ; PROTOCOLS ; DESIGN ; YIELD
英文摘要:

Background: Peripheral blood leukocytes (PBLs) are the main source of DNA in blood samples. A common protocol is to store buffy coat specimens for future DNA isolation, and such samples may remain in frozen storage for years. However, the currently available methods to maintain buffy coat specimens can be optimized for better quality and cost efficiency.

Study Design and Methods: Seventeen donors (aged 24-34 years) were enrolled in this study. Initially, five centrifugal speeds were chosen to examine the distribution of PBLs in the cell layer; the buffy coat was then harvested to evaluate the cell quantity and viability after storing at 4 degrees C for various times. Finally, the buffy coat was isolated, snap frozen, and kept at -80 degrees C for 1 hour, 1 week, or 4 weeks until the DNA was extracted. Agarose gel electrophoresis and multiplex PCR were used to verify DNA integrity and amplifiability.

Results: There was a linear positive correlation between the amount of fresh PBLs and the DNA yield. At least 70% of PBLs were collected in the uppermost 40% of the cellular material when the centrifugal force was over 800 g compared to 400 g. Storing blood at 4 degrees C for no more than 24 hours did not have an effect on the amount of PBLs or their viability. In addition, the amount of extracted DNA was decreased in the frozen buffy coat that was stored for more than 7 days, though the DNA quality was acceptable.

Conclusion: DNA should be extracted from fresh buffy coat samples as soon as possible after collection, especially for very important samples. Retaining the upper 40% of the cell pack of blood instead of whole blood could improve the storage efficiency of biobanking such samples. Our study provides a recommended option for blood collection and processing protocols in biobanking.

语种: 英语
WOS记录号: WOS:000349399300005
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/57550
Appears in Collections:北京大学第一临床医学院_期刊论文

Files in This Item:

There are no files associated with this item.


作者单位: 1.Key Lab Female Reprod Endocrine Related Dis, Shanghai, Peoples R China
2.Beijing Univ, Hosp 1, Beijing 100871, Peoples R China
3.Fudan Univ, Obstet & Gynecol Hosp, Shanghai 200011, Peoples R China

Recommended Citation:
Zhou, Jiayi,Wu, Jing,Zhao, Xiaoyan,et al. Biostorage and Quality Control for Human Peripheral Blood Leukocytes[J]. BIOPRESERVATION AND BIOBANKING,2015,13(1):13-19.
Service
Recommend this item
Sava as my favorate item
Show this item's statistics
Export Endnote File
Google Scholar
Similar articles in Google Scholar
[Zhou, Jiayi]'s Articles
[Wu, Jing]'s Articles
[Zhao, Xiaoyan]'s Articles
CSDL cross search
Similar articles in CSDL Cross Search
[Zhou, Jiayi]‘s Articles
[Wu, Jing]‘s Articles
[Zhao, Xiaoyan]‘s Articles
Related Copyright Policies
Null
Social Bookmarking
Add to CiteULike Add to Connotea Add to Del.icio.us Add to Digg Add to Reddit
所有评论 (0)
暂无评论
 
评注功能仅针对注册用户开放,请您登录
您对该条目有什么异议,请填写以下表单,管理员会尽快联系您。
内 容:
Email:  *
单位:
验证码:   刷新
您在IR的使用过程中有什么好的想法或者建议可以反馈给我们。
标 题:
 *
内 容:
Email:  *
验证码:   刷新

Items in IR are protected by copyright, with all rights reserved, unless otherwise indicated.

 

 

Valid XHTML 1.0!
Copyright © 2007-2017  北京大学医学部 - Feedback
Powered by CSpace