摘要
目的探讨人类活体胎儿血循环中的造血干/祖细胞(HSPC)表面标记物CD133、CD34、CD38随孕龄变化的规律.方法通过B超引导下脐静脉穿刺术等方法获得妊娠12～41周活体胎儿血106例,应用双色免疫荧光标记法分别标记单个核细胞CD34、CD133抗原和CD34、CD38抗原,流式细胞仪检测抗原表达.结果所有在B超引导下胎儿标本取样术后母胎未出现严重并发症,CD34+细胞/有核细胞、CD38-细胞/CD34+细胞、CD133+细胞/有核细胞、CD133+细胞/CD34+细胞含量均随孕周增加而降低,且与孕周呈直线负相关关系,其变化范围分别为4.21%～0.04%、58.5%～10.7%、3.69%～0.31%、87.6%～48.5%.结论越早期的胎儿循环血HSPC在免疫表型上越原始,这可能是宫内基因治疗理想的靶细胞.
Objective To investigate the dynamic development of fetal circulating hematopoietic stem/progenitor cell （HSPC） surface antigen CD133, CD34 and CD38 in on-going normal pregnancy. Methods 0.2 ～ 2.0 ml fetal blood samples were obtained from 106 human fetuses between the gestational age of postmenstrual weeks 12 and 41, 96 being collected by ultrasound-gnided percutaneous cordocentesis, 3 by ultrasound-guided selective feticide, and 7 being collected after birth. The nucleated cells （NCs） were separated, and the cell surface antigen CD34, CD38, and CD133 were labeled by bicolor immunofluorescence technique. Fluorescence activated cell sorting （FACS） was performed to analyze the concentration of CD34^＋/NC, CD38^-/CD34^＋, CD133^＋/NC, and CD133^＋/CD34^＋ cell respectively. Results All operations were uneventful for the fetuses and mothers. The proportions of CD34^＋/NC, CD38^-/CD34^＋ cells, CD133^＋/NC, and CD133^＋/CD34^＋ cells dramatically decreased when the gestational age advanced from week 12 to week 41. CD34^＋ cell/NC proportion decreased from 4.21% to 0.04% （r^2 =0.90, χ^-±s = 1.54% ±0.54% ）with the peak at gestational week 12 and a the second peak at the week 20. （2） The percentage of CD38^-/CD34^＋ cells decreased from 58.5% to 10.7% （ χ^- ± s = 28.66% ± 9.88% ） with advancing gestational age （r^2 =0.82） with the peak at gestational week 12 and a second peak at week 22. （3） The percentage of CD133^＋ cells among CD34^＋ cells decreased with advancing gestational age, from 87. 6% to 48.5% （ χ^-± s = 63.5% ± 11.4% ） . （4） CD133^＋ cells/NC concentration decreased with advancing gestational age, from 3.69% to 0.31% （χ^-± s = 1.40% ± 0.86% ） with the peak time at gestational week 12. , and showed a negative linear correlation with the gestational age （r^2 = 0.76）. Conclusion The immunophenotype of normal fetal circulating HSPC changes along with the gestational age in on-going pregnancy. The earlier the time, the more primitive the immunophenoptype of the fetal circulating HSPCs. The more primitive fetal circulating HSPCs may be the ideal target for in utero gene therapy.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2005年第28期1974-1977,共4页
National Medical Journal of China
基金
国家自然科学基金资助项目(30300373)
广东省科技攻关基金资助项目(2003C34220)
关键词
活体胎儿
循环血
造血干
/祖细胞
免疫表型
Fetus
Hematopoietic stem/Progenitor cell （HSPC）
Immunophenoptype
