| A study was carried out to develop media for the in vitro expansion of human and murine neural stem cells. Issues studied included the evaluation of basal media, the optimization of glucose, glutamine, hormone, amino acid, and buffer concentrations, and supplementation with growth factors, albumin, lipids, and trace elements. Based on these studies, two new media, PPRF.h2 and PPRF.m2, were developed for human and murine cells respectively.; The media were tested against a murine medium (NMSCM, published) and a human medium (NHSCM, proprietary) developed by NeuroSpheres Ltd. In static culture the human cells reached a maximum density in PPRF.h2 which was 10% higher than in NHSCM. In static culture the maximum murine cell density in PPRF.m2 was 75% greater than in NHSCM and 655% greater than in NMSCM. In stirred culture they were 53.9% and 503% greater respectively. Triple staining of the murine cells after growing in PPRF.m2 revealed that they retained their ability to give rise to neurons, astrocytes, and oligodendrocytes. |
