Full Text:


영남의대학술지 Vol.24_No.2 Suppl. P.S555-561, Dec. 2007

Original Article

자가혈청이 배양연골세포의 부착과 증식에 미치는 효과*

The Effects of Autologous Serum on the Attachment and Proliferation of Human Primary Chondrocyte Culture

영남대학교 의과대학 성형외과학교실
책임저자:정재호, 대구광역시 남구 대명동 317-1, 영남대학교 의과대학 성형외과학교실
Tel: (053) 620-3482, Fax: (053) 626-0705
E-mail: jhjeong@med.yu.ac.kr

December 30, 2007


Background:Culture method for expanding chondrocytes ex vivo is an important procedure in cartilage tissue engineering. In most laboratories related to tissue engineering, fetal bovine serum is widely used as supplement. However, the chondrocytes grown in a medium containing fetal bovine serum may cause infectious viral diseases such as Creutzfeldt-Jacob disease and/or unfavorable immune reaction to bovine proteins. As a way out of these problems, we examined whether a patient's autologous serum could support the growth and attachment of his/her chondrocytes.
Materials and Methods:Chondrocytes were isolated from microtia patients (age between 5 to 12) by enzymatic digestion and cultured in a medium supplemented with 10% autologous serum, 10% fetal bovine serum (FBS) and 10% banked allogenic serum respectively. Proliferation and attachment rate were assessed by Trypan blue staining and MTT assay. Attachment rate was checked at 6, 12, 24, 36, 48 hours after plating of cells and counting was done with hemocytometer after trypan blue exclusion. Proliferation rate was checked at 1, 3, 5, 7, 9 days after plating of cells and measurement was done with cell counting and MTT assay.
Results:As a result, chondrocytes which were cultured in the medium supplemented with 10% autologous serum, represented higher rate of both proliferation and attachment, which is comparable to the chondrocytes in the culture supplemented with 10% FBS. But, chondrocytes in the culture supplemented with 10% banked allogenic serum showed lower rate of proliferation and attachment.
Conclusion:The beneficial effect of autologous serum which has been confirmed in this study is another important progress for clinical application of tissue engineering. The possibility of banked allogenic serum is still remained. In this study, we used banked allogenic serum which containing anti-coagulants and this component may have affected on the result. Fresh allogenic serum should be utilized for next step of experiment.

*본 연구는 한국과학재단 목적기초연구(과제번호: R05-2000-000-00171-0) 지원으로 수행되었음.

Key Words: Cartilage tissue engineering, Autologous serum, Chondrocyte culture


1. Maurer HR. Towards chemically defined serum-free media for mammalian cell culture. Animal cell culture 1992;2:15-19.

2. Verbruggen G, Verdonk R, Veys EM, Van Daele P, De Smet P, Van den Abbeele K, et al. Human meniscal proteoglycan metabolism in long-term tissue culture. Knee Surg Sports Traumatol Arthrosc. 1996;4(1):57-63.

3. Lennon DP, Haynesworth SE, Young RG, Dennis JE, CaplanAI. A chemically defined medium supports in vitro proliferation and maintains the osteochondral potential of rat marrow-derived mesenchymal stem cells. Exp Cell Res 1995 Jul;219(1):211-22.

4. Nijweide PJ, Burger EH. Mechanisms of bone formation in vitro. In The osteoblast(Ed. B. K. Hall), Telford Press, Caldwell, New Jersey. 1990;1:303.

5. Gruber R, Sittinger M, Bujia J. In vitro cultivation of human chondrocytes using autologous human serum supplemented culture medium: minimizing possible risk of infection with pathogens of prion diseases. Laryngor- hinootologie 1996 Feb;75(7):105-8.

6. Hankey DP, McCabe RE, Doherty MJ, Nolan PC, McAlinden MG, Nelson J, et al. Enhancement of human osteoblast proliferation and phenotypic expression when cultured in human serum. Acta Orthop Scand 2001 Aug;72(4):395-403.

7. Choi YH, Lee SJ, Nguyen P, Jang JS, Lee J, Wu ML, et al. Regulation of cyclin D1 by calpain protease. J Biol Chem 1997 Nov;7:272 (45):28479-84.