A method for isolation and cultivation of adult Schwann cells for nerve conduit
Azim Hedayatpour1, Aligholi Sobhani, Vahid Bayati
1Department of Anatomy, Faculty of Medicine, Medical Sciences/University of Tehran, Tehran, Iran.
Background:
It has been found that one of the methods to repair peripheral nervous system or even central nervous system injury is to use Schwann cells as nerve regeneration promoters. Therefore, it seems necessary to look for a way to obtain activated Schwann cells, with a sufficient amount of numbers and purity, in a short time for clinical applications. However, the previous methods using mitogens are not much clinically acceptable, and other methods that do not require mitogens, fail to isolate adult Schwann cells effectively or require a long period of time.
Methods:
In this study, Schwann cells were isolated from predegenerated sciatic nerves of adult rat (one to three nerves per primary culture) and subcultivated two times in a week with the 10%fetal bovine serum supplementation. Thereafter, Dulbecco's Modified Eagle's Medium media supplemented with 10%, 5%, 2.5%, 1.25%, and 0.625% fetal bovine serum were employed to determine their influence on the density and purity of Schwann cells after a 10-day period of cultivation.
Results:
The concentrations of fetal bovine serum less than 10% immediately stimulated some morphological changes to happen in Schwann cells but not fibroblasts. Finally, Schwann cells acquired their normal shape on day 6 when fibroblasts just began to alter and die.
Conclusion:
Our results demonstrated that total cell density was highly significant (P<0.05) in the medium supplemented with 10% fetal bovine serum (950 cells/mm(1)) while purity was significant (P<0.05) in the medium supplemented with 2.5% fetal bovine serum (97%) in comparison with other concentrations of fetal bovine serum.
