Document Detail


A new bone banking technique to maintain osteoblast viability in frozen human iliac cancellous bone.
MedLine Citation:
PMID:  12237093     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The aim of this study was to develop a new cryopreservation technique to maintain the osteoblast viability in frozen iliac bone and to prove cell viability using cell culture techniques. Human iliac cancellous bones were frozen with and without 10% Me(2)SO at -80 degrees C. The tubes were kept in a -80 degrees C freezer for at least 2 days. After the storage period, the frozen bone was thawed by placing the tube in a 37 degrees C water bath. A serial enzymatic digestion technique using 0.2% collagenase was employed to isolate osteoblast-like cells from the bone. The cells that were released were inoculated into tissue culture flasks containing DMEM supplemented with 10% FCS. They were incubated at 37 degrees C in a humidified atmosphere of 95% air and 5% CO(2). Cells of the second passage were plated at a density of 5 x 10(3)cells/cm(2) in a 24-well plate and used for characterization. For characterization, WST-1 assay, determination of alkaline phosphatase, Type I collagen assay, osteocalcin assay, and von Kossa staining were used. The assays were performed at 3, 6, 9, and 12 days after plating the cells. Based on the results of this study, we conclude that the osteoblast-like cells in the frozen bone can survive, only when the bone is frozen with cryoprotectants to prevent injury during freezing and thawing.
Authors:
Jung-Hwan Oh; Joachim E Zöller; Alexander Kübler
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Publication Detail:
Type:  In Vitro; Journal Article    
Journal Detail:
Title:  Cryobiology     Volume:  44     ISSN:  0011-2240     ISO Abbreviation:  Cryobiology     Publication Date:  2002 Jun 
Date Detail:
Created Date:  2002-09-18     Completed Date:  2003-05-27     Revised Date:  2009-11-03    
Medline Journal Info:
Nlm Unique ID:  0006252     Medline TA:  Cryobiology     Country:  United States    
Other Details:
Languages:  eng     Pagination:  279-87     Citation Subset:  IM    
Affiliation:
Department of Oral and Maxillofacial Surgery, University of Cologne, Kerpener Str. 62, 50937, Cologne, Germany.
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MeSH Terms
Descriptor/Qualifier:
Alkaline Phosphatase / metabolism
Bone Matrix / cytology,  metabolism
Cell Division
Cell Survival
Cells, Cultured
Collagen Type I / metabolism
Cryopreservation / methods*
Cryoprotective Agents
Dimethyl Sulfoxide
Humans
Ilium* / cytology,  metabolism
Osteoblasts* / cytology,  metabolism
Osteocalcin / metabolism
Tissue Banks
Tissue Preservation / methods*
Chemical
Reg. No./Substance:
0/Collagen Type I; 0/Cryoprotective Agents; 104982-03-8/Osteocalcin; 67-68-5/Dimethyl Sulfoxide; EC 3.1.3.1/Alkaline Phosphatase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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