Pediatric Urology and Regenerative Medicine Research Center, Section of Tissue Engineering and Stem Cells Therapy, Children's Hospital Medical Center, Tehran University of Medical Sciences, Tehran, Iran.
Int Braz J Urol. 2018 May-Jun;44(3):608-616. doi: 10.1590/S1677-5538.IBJU.2017.0324.
To identify the fetal stem cell (FSC) response to maternal renal injury with emphasis on renal integrity improvement and Y chromosome detection in damaged maternal kidney.
Eight non-green fluorescent protein (GFP) transgenic Sprague- Dawley rats were mated with GFP-positive transgenic male rats. Renal damage was induced on the right kidney at gestational day 11. The same procedure was performed in eight non-pregnant rats as control group. Three months after delivery, right nephrectomy was performed in order to evaluate the injured kidney. The fresh perfused kidneys were stained with anti-GFP antibody. Polymerase chain reaction (PCR) assay was also performed for the Y chromosome detection. Cell culture was performed to detect the GFP-positive cells. Technetium-99m-DMSA renal scan and single-photon emission computed tomography (SPECT) were performed after renal damage induction and 3 months later to evaluate the improvement of renal integrity.
The presence of FSCs was confirmed by immune histochemical staining as well as immunofluorescent imaging of the damaged part. Gradient PCR of female rat purified DNA demonstrated the presence of Y-chromosome in the damaged maternal kidney. Moreover, the culture of kidney cells showed GPF- positive cells by immunofluorescence microscopy. The acute renal scar was repaired and the integrity of damaged kidney reached to near normal levels in experimental group as shown in DMSA scan. However, no significant improvement was observed in control group.
FSC seems to be the main mechanism in repairing of the maternal renal injury during pregnancy as indicated by Y chromosome and GFP-positive cells in the sub-cultured medium.
确定胎儿干细胞(FSC)对母体肾损伤的反应,重点关注受损母体肾脏中肾完整性的改善和 Y 染色体的检测。
将 8 只非绿色荧光蛋白(GFP)转基因 Sprague-Dawley 大鼠与 GFP 阳性转基因雄性大鼠交配。在妊娠第 11 天对右侧肾脏进行肾损伤。对 8 只未怀孕的大鼠进行相同的操作作为对照组。分娩 3 个月后,行右肾切除术以评估损伤的肾脏。用抗 GFP 抗体对新鲜灌注的肾脏进行染色。还进行了聚合酶链反应(PCR)检测以检测 Y 染色体。进行细胞培养以检测 GFP 阳性细胞。在肾损伤诱导后和 3 个月后进行锝-99m-DMSA 肾扫描和单光子发射计算机断层扫描(SPECT),以评估肾完整性的改善。
免疫组织化学染色和损伤部位的免疫荧光成像证实了 FSCs 的存在。对雌性大鼠纯化 DNA 的梯度 PCR 显示受损母体肾脏中存在 Y 染色体。此外,通过免疫荧光显微镜观察,培养的肾细胞显示 GFP 阳性细胞。在实验组中,急性肾瘢痕得到修复,损伤肾脏的完整性接近正常水平,DMSA 扫描显示;然而,对照组未见明显改善。
FSC 似乎是妊娠期间修复母体肾损伤的主要机制,这表明在亚培养培养基中存在 Y 染色体和 GFP 阳性细胞。