Culture of rat cerebral oligodendrocytes in a serum-free, chemically defined medium

Abstract

Kew'ords: oligodendrocyte--primary culture--serum-free medium--chemically defined medium--brain cells--rat brain Oligodendrocytes were isolated from the cerebra of young rats (5-10 days old) by trypsinization of the tissue followed by cell separation on Percoll gradients. The isolation was carried out in physiological, isotonic media. The cell yield was 2-4 × 106 cells per brain; the plating efficiency was >/70%. Isolated cells were seeded on poly-L-lysine-coated culture dishes and maintained in a serum-free, chemically defined medium for at least 30 days. After 10 days in culture 67+10% of the surviving cells were oligodendrocytes, as judged by immunocytochemical and morphological criteria, whereas most of the other cells reacted positively with antiserum against glial fibrillary acidic protein. The expression of typical oligodendrocyte markers (2':3'-cyclic-nucleotide 3'-phosphodiesterase, galactocerebrosides and myelin basic protein) was greatly enhanced under these serum-free conditions as compared with cultures in serum-containing medium. The antigenic markers (galactocerebrosides, myelin basic protein) were absent in the freshly isolated cells but could be detected after 3 days in culture by immunocytochemistry. The activity of 2':3'-cyclic-nucleotide 3'-phosphodiesterase increased from 75 nmol min 1 mg 1 protein on day 4 to 400 nmol min-1 mg 1 protein on day 14 in culture.

Acetoacetate and glucose as substrates for lipid synthesis by rat brain oligodendrocytes and astrocytes in serum-free culture

“…In the present study we have investigated the contributions of glucose and acetoacetate to lipogenesis and cholesterogenesis in a primary culture system of oligodendrocytes and astrocytes maintained in a serum-free medium [18]. The metabolic processes occurring in oligodendrocytes and astrocytes could be distinguished by developing a method for complement-mediated cytolysis of the oligodendrocyte popula~on in these mixed glial cultures.…”

Growth requirements in vitro of oligodendrocyte cell lines and neonatal rat brain oligodendrocytes.

“…A different CDM for culture of isolated oligodendrocytes from 5-to 10-day-old rat cerebra was described by Koper et al. (38). Cells were maintained for 30 days in this CDM, which is very complex and contains 19 supplements, including the 4 supplements in 01 medium.…”

Tumour necrosis factor- , interferon- and interferon-beta exert antiviral activity in nervous tissue cells

“…Glial cell cultures were prepared as previously described (Koper et al, 1984). Briefly, cerebra of 7-day-old Wistar rats were minced and incubated with trypsin (45 rain at 37 °C).…”