Primary chondrocyte culture in medium supplemented with autologous serum. Applicability in endoscopic treatment of vesico-ureteral reflux
Article published in Urologia Polska 2004/57/2.

authors

Tomasz Drewa ^1,2, Zbigniew Wolski ¹, Przemysław Gałązka ², Artur Czaplewski ², Alina Woźniak ², Jan Sir ³

¹ Katedra i Klinika Urologii Akademii Medycznej w Bydgoszczy
Kierownik kliniki: prof. dr hab. Zbigniew Wolski
² Katedra i Zakład Biologii Medycznej Akademii Medycznej
Kierownik zakładu: prof. dr hab. Gerard Drewa
³ Zakład Patologii Nowotworów, Regionalne Centrum Onkologii w Bydgoszczy
Kierownik zakładu: lek. med. Jan Sir

keywords

autologous serum, chondrocytes transplantation, vesico-ureteral reflux

summary

introduction

Transplanted autologous chondrocytes (ACT) can be used in endoscopic treatment of vesico-ureteral reflux. Fetal bovine serum (FBS) is still a gold standard for media supplementation in many laboratories. Prion and virus diseases are a danger for patients treated with ACT if fetal bovine serum FBS is used.

aim

The aim of this study was to establish the utility of media for chondrocytes culture supplemented with Autologous Serum (AS) or serum substitute Ultroser G (UG) instead of FBS.

material and methods

Cartilage specimens were obtained form knee joints of 5 young rabbits and digested during 10 hours with collagenase II and hyaluronidase. Autologous rabbit serum (AS) was prepared. Ham`s F-12 medium was supplemented with 10% (v/v) of FBS (Sigma) or 10% (v/v) AS. Ultroser G (UG, Gibco) was added in concentration of 2% (v/v). After 14 days living cells were counted using trypan blue.

results

It was not possible to obtain primary chondrocytes cultures using medium supplemented with Ultroser G. The mean total number of cells in primary cultures supplemented with 10% FBS was 5,81x106 while mean number of cells growing in medium with 10% AS was 7,27x106. Medium supplemented with AS in concentration of 10% shown the best proliferation effect on chondrocytes cultures (PRCC). There were no differences in cells morphology between cells growing in medium supplemented with 10% FBS and 10% AS.

conclusions

Our preliminary results confirm the possibility of using autologous serum (AS) for establishing the primary chondrocytes cultures.