Khosrotehrani K, Reyes R R, Johnson K L, Freeman R B, Salomon R N, Peter I, Stroh H, Guégan S, Bianchi D W
Division of Genetics, Department of Pediatrics, Tufts-New England Medical Center, Boston, MA 02111, USA.
Hum Reprod. 2007 Mar;22(3):654-61. doi: 10.1093/humrep/del426. Epub 2006 Oct 30.
In humans, fetal microchimeric cells transferred to maternal tissues during pregnancy can adopt a hepatocyte phenotype. Our objective was to determine whether fetal cells participate in the response to specific murine post-partum hepatic injuries.
Wild-type female mice were bred to males transgenic for the enhanced green fluorescent protein (GFP) (n = 42). Following delivery, we created models of chemical or surgical injury with carbon tetrachloride (CCl(4)) injection or by performing partial hepatectomy. Liver injury was assessed histologically. Fetal cells in maternal liver were detected and measured by real-time PCR amplification of the gfp transgene and by immunofluorescence using anti-GFP antibodies.
PCR results showed that in chemical but not surgical injury, fetal GFP+ cells were detectable in maternal liver and spleen and that fetal cell presence was significantly increased over time following injury (4 versus 8 weeks, P = 0.006 for liver and P = 0.0006 for spleen). In some animals, following chemical injury, GFP+ cells were detected by immunofluorescence.
The results of this preliminary study suggest that specific types of injury may elicit different fetal cell responses in maternal organs. There is a significant effect of time on fetal cell presence in liver and spleen. Furthermore, real-time PCR amplification is more sensitive than immunofluorescence for the detection of microchimeric fetal cells.
在人类中,孕期转移至母体组织的胎儿微嵌合细胞可呈现肝细胞表型。我们的目的是确定胎儿细胞是否参与对特定小鼠产后肝损伤的反应。
将野生型雌性小鼠与增强型绿色荧光蛋白(GFP)转基因雄性小鼠交配(n = 42)。分娩后,我们通过注射四氯化碳(CCl₄)或进行部分肝切除术创建化学或手术损伤模型。通过组织学评估肝损伤。通过对gfp转基因进行实时PCR扩增以及使用抗GFP抗体进行免疫荧光检测和测量母体肝脏中的胎儿细胞。
PCR结果显示,在化学损伤而非手术损伤中,可在母体肝脏和脾脏中检测到胎儿GFP⁺细胞,并且损伤后随着时间推移胎儿细胞的存在显著增加(4周与8周相比,肝脏P = 0.006,脾脏P = 0.0006)。在一些动物中,化学损伤后通过免疫荧光检测到GFP⁺细胞。
这项初步研究的结果表明,特定类型的损伤可能在母体器官中引发不同的胎儿细胞反应。时间对肝脏和脾脏中胎儿细胞的存在有显著影响。此外,实时PCR扩增在检测微嵌合胎儿细胞方面比免疫荧光更敏感。