A Study on the Formation of Organotypic Spheroids from Early Human Fetal Brain

Jung, Hee Won; Han, Jong Woo; Kim, Jong Soo; Chung, Chun Kee; Jeon, Sang Ryong; Choi, Kil Soo

Abstract

The present study investigated whether organotypic spheroids derived from human fetal brain tissue, cultured at agarose-overlay media, may serve as an optimal invitro model for multidisciplinary studies in human neurobiology, particularly in the fields of tumor invasiveness and its biochemical mechanism, using light microscopy, electron microscopy and immunohistochemical staining. Eight fetal brain tissues of 8-9 weeks of gestation were minced and explanted into agarose-coated culture wells. After three to five days these human fetal brain tissue fragments emerged as spheroids and could be maintained as organotypic spheroids for up to seven weeks. Light and electron microscopic studies of sphereoids demonstrated that most cells were poorly differentiated and there were no definite mature neurons or glial cells after enough culture time, but some cells showed certain evidence suggestive of differentiation to neurons or glial cells. Immunohistochemical staining for glial fibrillary acidic protein(GFAP) and neur on specific enolase(NSE) demonstrated that NSE-positive cells were oval or spherical cells containing abundant cytoplasm and GFAP-positive cells were fibrillary cytoplasma-containing cells which showed some evidence suggestive of differentiation to glial cells by light microscopy. In future, adding some modifications in culture, this organotypic spheroids derived from the human fetal brain may serve as an optimal in vitro model for neurobiology especially in the field of studies on tumor invasiveness through co-culture with microtumor spheroids.