Comparison of cancellous bone-derived cell proliferation in autologous human and fetal bovine serum

Abstract

Conventionally, culture medium is supplemented with fetal bovine serum (FBS): such serum presents potential risks of foreign protein contamination and transmission of viral or prion-related disease if used in culture of cells intended for human reimplantation. As it has been suggested that a composite of cultured human cancellous bone-derived cells and a bone graft substitute may present a solution to the well-recognized complications and limited availability associated with harvest of fresh bone graft, this study aimed to compare the proliferative response of human cancellous bone-derived cells supplemented with FBS or autologous human serum (AHS) to determine whether AHS is a practical alternative. Explant cultures were established using greater trochanter trabecular bone from 10 consenting patients (aged 57-84) undergoing total hip arthroplasty. At the same time, serum was harvested. The cells were characterized by alkaline phosphatase expression and by in vitro mineralization in enhanced medium. At confluence, cells were aliquoted into multiwell plates and grown for 9 days in medium supplemented with 5%, 10%, 15%, or 20% AHS or 10% FBS. Proliferative response was determined by a crystal violet dye binding assay. There was no significant difference between proliferation in 5% AHS and 10% FBS. However, 10%, 15%, and 20% AHS all produced significantly greater proliferation than 10% FBS. The proliferative response was dose related. FBS is said to be rich in growth and attachment factors, which is why it is widely used in tissue culture. These results suggest that species specificity, even when using adult serum, outweighs these advantages. It should therefore be considered as a prerequisite for any program involving reimplantation of cultured human cells. Clinical trials of cultured human cancellous bone-derived cells have now begun.