Characterization of an established human malignant glioma cell line: LN-18

Abstract

A human malignant glioma cell line, LN-18, has been established in monolayer culture and subcultured for more than 115 passages. LN-18 cells grow in vitro as bipolar or stellate cells with pleomorphic nuclei, have a doubling time of about 72 h and a plating efficiency of 3%. The glial nature of these cells has been assessed by ultrastructural examination. The synthesis of glial fibrillary acidic and S-100 proteins could not be demonstrated, although the initial biopsy tissue and the early cultures were positive for the former. The presence of Ia-like antigens on the surface of these cells was demonstrated using allo and xeno antisera. LN-18 cells were also shown to synthesize large quantities of fibronectin. The injection of LN-18 cells into nude mice induced the formation of solid tumor masses that could be retransplanted every 3 weeks and showed a morphology comparable to that of the initial biopsy. Karyotype analysis revealed the presence of three marker chromosomes, constantly present before and after hetero-transplantation.

Renin and angiotensinogen expression and functions in growth and apoptosis of human glioblastoma

“…Having established the presence of most of the RAS components in human glioblastoma, we addressed its role in tumour cells using the human LN18 and LNZ308 glioblastoma cell lines (Diserens et al, 1981;Egidy et al, 2000;Juillerat-Jeanneret et al, 2000). Both cell lines expressed renin, renin receptor, AGT and AT 2 mRNAs using RT -PCR, while only LNZ308 expressed AT 1 mRNA ( Figure 3A).…”

“…The AT 2 antagonist PD123319 was purchased from Sigma. LN18 and LNZ308 human glioblastoma cells (a kind gift of AC Diserens, CHUV, Lausanne, Switzerland; Diserens et al, 1981) were grown in DMEM medium containing 4.5 g l À1 glucose and 5% FCS. Cells were split in 48-well plates and cultured for 1 -3 days until confluence was reached.…”

INK4a/Arf is required for suppression of EGFR/ΔEGFR(2-7)-dependent ERK activation in mouse astrocytes and glioma

“…The brain was extracted, formalin fixed, and paraffin embedded for histology. Macroscopically visible tumors were put in culture to derive cell lines as described previously (Diserens et al, 1981). Immunohistochemistry was performed under standard conditions using a heat epitope retrieval method in citrate buffer (pH 6.0; 3 min in pressure cooker) with the exception for GFAP (anti-EGFR, 1 : 200, sc-03 1005, Santa Cruz, CA, USA; anti-GFAP, 1 : 200, Z0334, Dako, Glostrup, Denmark; anti-cyclin D1/2, 1 : 500, 05-362, Upstate, Lake Placid, NY, USA).…”

Regulation of Aminopeptidase A in Human Brain Tumor Vasculature: Evidence for a Role of Transforming Growth Factor-β

“…This antibody did not recognize any structure in our conditions (not shown). Low intracellular levels of active renin (ranging from 8.6 to 60.9 pg renin/mg protein) could be detected in the human glioblastoma cell lines LN18, LN229, LN319, and LN428 (Diserens et al, 1981), but no renin could be detected in culture supernatants. The four glioblastoma cell lines did not express ACE activity and did not inhibit the activity of purified plasma ACE (not shown).…”

“…Three to four million glioblastoma cells (Diserens et al, 1981) were washed with PBS, detached from the plate with a rubber policeman, and extracted by freeze/ thawing (3 times) in 1 ml of 0.1 M phosphate, 0.3 M NaCl, pH 8.0. Extracts were centrifuged, and the concentration of renin was measured in supernatants using an IRMA-renin determination kit (Sanofi-Pasteur, BioRad SDP, Reinach, Switzerland).…”