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In vivo establishment of T98G human glioblastoma
M Rubenstein1, M Shaw, Y Mirochnik
1Department of Urology, Rush Presbyterian St. Lukes Medical Center, Chicago, IL, USA. DrMarv@prodigy.net
Abstract:
Human derived T98G glioblastoma has long been utilized as an in vitro model for epidermal growth factor receptor (EGFR)-mediated growth regulation. Recently, T98G has been employed to develop new types of therapy directed at limiting EGFR expression such as by administration of antisense oligonucleotides directed against EGFR encoding mRNA. A major limitation to extending this model for in vivo application is that T98G implanted s.c. or intracerebrally has been reported not to grow in nude mice. In an effort to extend this model to permit in vivo studies, we evaluated the use of Matrigel and orthotopic (intracranial) implantation techniques. When equal volumes of Matrigel were mixed with T98G cell suspensions, tumors developed at both flank and orthotopic locations. Four groups of nude mice were inoculated into the flanks with either 10(5), 10(6), 4 x 10(6) or 10(7) T98G cells in a 150 microliters total volume with Matrigel. In 1/5, 3/5, 1/5 and 1/3 mice receiving 10(5), 10(6), 4 x 10(6) and 10(7) cells, respectively, tumors developed 11, 15, 15 and 15 weeks, respectively, following inoculation. Out of 4 mice inoculated orthotopically (intracranially into the frontal lobe) with only 4 x 10(4) cells and Matrigel, 2 developed tumors. However, all mice (4/4) inoculated orthotopically with 4 x 10(5) cells in a 10 microliters total volume with Matrigel developed tumors. Two were identified histologically following a scheduled sacrifice at 36 and 60 days and two more at 103 and 118 days after sacrifice following abnormal behavior. The best tumor establishment efficacy combined orthotopic implantation of 4 x 10(5) T98G cells with Matrigel. These techniques permit the use of T98G glioblastoma as an in vivo model for new forms of therapy.
Insights
This study shows that combining Matrigel with orthotopic implantation of T98G glioblastoma cells enables in vivo tumor development. These methods allow T98G glioblastoma to be used as an in vivo model for novel therapies.
Area of Science:
- Neuro-oncology
- Cancer biology
- Biotechnology
Background:
- T98G glioblastoma is a human cell line used as an in vitro model for epidermal growth factor receptor (EGFR)-mediated growth.
- Previous attempts to use T98G cells for in vivo studies in nude mice were limited due to poor tumor formation.
- New therapies targeting EGFR expression, such as antisense oligonucleotides, require viable in vivo models for testing.
Purpose of the Study:
- To establish a reliable in vivo model using T98G glioblastoma cells for preclinical therapeutic studies.
- To evaluate the efficacy of Matrigel and orthotopic implantation techniques for T98G tumor development in nude mice.
Main Methods:
- T98G glioblastoma cells were mixed with Matrigel and implanted either subcutaneously (flank) or orthotopically (intracranially) into nude mice.
- Various cell concentrations were tested for flank and orthotopic implantations.
- Tumor development and efficacy were monitored over time.
Main Results:
- Matrigel combined with orthotopic implantation of T98G cells significantly improved tumor establishment.
- Orthotopic implantation of 4 x 10^5 T98G cells with Matrigel resulted in 100% tumor formation in nude mice.
- Subcutaneous implantation showed lower tumor take rates and longer latency periods compared to orthotopic implantation.
Conclusions:
- The combination of Matrigel and orthotopic implantation provides an effective method for establishing T98G glioblastoma in vivo.
- This optimized model facilitates the in vivo evaluation of novel therapies targeting glioblastoma, particularly those related to EGFR signaling.