A new bone banking technique to maintain osteoblast viability in frozen human iliac cancellous bone

Authors: Oh, J-H.; Zo¨ller, J.E.; Ku¨bler, A.

Source: Cryobiology, Volume 44, Number 3, June 2002 , pp. 279-287(9)

Publisher: Academic Press

Buy & download fulltext article:

The full text article is not available for purchase.

The publisher only permits individual articles to be downloaded by subscribers.


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%Me2SO at -80 °C. The tubes were kept in a -80 °C freezer for at least 2 days. After the storage period, the frozen bone was thawed by placing the tube in a 37 °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 °C in a humidified atmosphere of 95% air and 5%CO2. Cells of the second passage were plated at a density of5×103cells/cm2 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.

© 2002 Elsevier Science (USA)

Document Type: Research Article

DOI: http://dx.doi.org/10.1016/S0011-2240(02)00034-2

Publication date: June 1, 2002

Related content



Free Content
Free content
New Content
New content
Open Access Content
Open access content
Subscribed Content
Subscribed content
Free Trial Content
Free trial content

Text size:

A | A | A | A
Share this item with others: These icons link to social bookmarking sites where readers can share and discover new web pages. print icon Print this page