Key Laboratory of Health Ministry for Congenital Malformation, Shengjing Hospital, China Medical University, Shenyang, China.
Key Laboratory of Health Ministry for Congenital Malformation, Shengjing Hospital, China Medical University, Shenyang, China.
Cytotherapy. 2014 Mar;16(3):319-30. doi: 10.1016/j.jcyt.2013.10.005. Epub 2013 Dec 22.
In a previous study, we successfully devised a prenatal surgical approach and transplanted mesenchymal stromal cells (MSCs) to fetal rat spinal column to treat retinoic acid-induced neural tube defects in rat. Our results show that MSCs survived, migrated and differentiated into neural lineage cells. We intended to study various growth factor expressions in rat fetal spinal cords with spina bifida aperta after in utero MSC transplantation and the effect of in vivo growth factor introduction for prenatal spina bifida treatment.
Pregnant rats were treated with retinoic acid on embryonic day 10 and then received fetal surgery for MSC transplantation and/or lentiviral epidermal growth factor (EGF) injection on embryonic day 16; various growth factor expression in spinal cords from embryonic day 20 fetuses were analyzed by means of quantitative reverse transcriptase-polymerase chain reaction. Terminal deoxynucleotidyl transferase dUTP nick end labeling analysis was performed to observe spinal tissue apoptosis.
Growth factor expression was dysregulated in spinal cords with spina bifida. After MSC transplantation, we observed significantly increased expression of EGF, fibroblast growth factor (FGF)-8, FGF-2 and FGF-20 in the MSC transplantation group compared with blank injection; Furthermore, EGF expression positively correlated with surviving MSC amounts. Expression of other growth factors was not significantly different. In vivo EGF introduction reduced spinal tissue apoptosis.
Our results suggest that intrinsic EGF and FGF-2, FGF-8 and FGF-20 might affect the in vivo fate of transplanted MSCs in a fetal rat spina bifida model. In vivo EGF introduction together with MSC transplantation might serve as a new strategy for prenatal spina bifida treatment.
在之前的研究中，我们成功设计了一种产前手术方法，并将间充质基质细胞（MSCs）移植到胎儿大鼠脊柱中，以治疗大鼠中视黄酸诱导的神经管缺陷。我们的结果表明，MSCs 存活、迁移并分化为神经谱系细胞。我们旨在研究宫内 MSC 移植后伴有开放性脊柱裂的大鼠胎儿脊髓中的各种生长因子表达，并研究体内生长因子引入对产前脊柱裂治疗的影响。
妊娠大鼠于胚胎第 10 天接受视黄酸处理，然后于胚胎第 16 天进行胎儿手术进行 MSC 移植和/或慢病毒表皮生长因子（EGF）注射；通过定量逆转录聚合酶链反应分析胚胎第 20 天胎儿脊髓中的各种生长因子表达。末端脱氧核苷酸转移酶 dUTP 缺口末端标记分析观察脊髓组织凋亡。
脊柱裂脊髓中的生长因子表达失调。MSC 移植后，我们观察到 MSC 移植组 EGF、成纤维细胞生长因子（FGF）-8、FGF-2 和 FGF-20 的表达明显增加，与空白注射组相比；此外，EGF 表达与存活的 MSC 数量呈正相关。其他生长因子的表达没有明显差异。体内 EGF 引入可减少脊髓组织凋亡。
我们的结果表明，内源性 EGF 和 FGF-2、FGF-8 和 FGF-20 可能影响胎儿大鼠脊柱裂模型中移植 MSC 的体内命运。体内 EGF 引入联合 MSC 移植可能成为产前脊柱裂治疗的新策略。
