Rubenstein M, Shaw M, Mirochnik Y, Slobodskoy L, Glick R, Lichtor T, Chou P, Guinan P
Department of Urology, Rush Presbyterian St. Lukes Medical Center, Chicago, IL, USA.
Methods Find Exp Clin Pharmacol. 1999 Jul-Aug;21(6):391-3. doi: 10.1358/mf.1999.21.6.541918.
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.
人源T98G胶质母细胞瘤长期以来一直被用作表皮生长因子受体（EGFR）介导的生长调节的体外模型。最近，T98G已被用于开发旨在限制EGFR表达的新型疗法，例如通过给予针对EGFR编码mRNA的反义寡核苷酸。将该模型扩展用于体内应用的一个主要限制是，据报道，皮下或脑内植入的T98G在裸鼠中不会生长。为了扩展该模型以允许进行体内研究，我们评估了基质胶和原位（颅内）植入技术的使用。当将等体积的基质胶与T98G细胞悬液混合时，在双侧胁腹和原位位置均形成了肿瘤。将四组裸鼠的胁腹接种150微升总体积、含基质胶的10⁵、10⁶、4×10⁶或10⁷个T98G细胞。分别接受10⁵、10⁶、4×10⁶和10⁷个细胞的小鼠中，肿瘤分别在接种后11、15、15和15周形成。在仅接种4×10⁴个细胞和基质胶的4只原位（颅内额叶）接种的小鼠中，2只形成了肿瘤。然而，所有4只原位接种10微升总体积、含基质胶的4×10⁵个细胞的小鼠均形成了肿瘤。在预定于36天和60天处死后，通过组织学鉴定出2只肿瘤小鼠，在行为异常后处死的103天和118天后又鉴定出2只。最佳的肿瘤形成效果是将4×10⁵个T98G细胞与基质胶原位植入相结合。这些技术允许将T98G胶质母细胞瘤用作新型疗法的体内模型。
