سلول‌های جنینی در خون مادر: جنبه‌های بالینی و تکنیکی

نوع مقاله : مقاله مروری

نویسندگان

1 دانشجوی کارشناسی ارشد، مرکز تحقیقات بیماری‌های ارثی کودکان و گروه ژنتیک و بیولوژی مولکولی، دانشکده‌ی پزشکی و کمیته‌ی تحقیقات دانشجویی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

2 دانشیار، مرکز تحقیقات بیماری‌های ارثی کودکان و گروه ژنتیک و بیولوژی مولکولی، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

چکیده

روش‌های رایج برای به دست آوردن نمونه از جنین برای تشخیص پیش از تولد، برای جنین و مادر خطرناک می‌باشد. این موضوع سبب انجام تلاش‌های زیاد دانشمندان در زمینه‌ی به دست آوردن سلول‌های جنینی به روش‌های غیر تهاجمی شده است که ازآن جمله، به دست آوردن آن‌ها از خون محیطی مادران باردار قابل ذکر است. انواع سلول‌هایی که توسط بسیاری از دانشمندان در سر تا سر جهان بر روی آن‌ها در این زمینه مطالعه صورت گرفته است، شامل لوکوسیت‌های جنینی به ویژه لنفوسیت‌ها و گرانولوسیت‌ها، گلبول‌های قرمز هسته‌دار جنینی و سلول‌های تروفوبلاست می‌باشد. بسیاری از مطالعات بر روی گلبول‌های قرمز هسته‌دار جنینی صورت گرفته است که از علل این توجه می‌توان به میزان بالای این سلول‌ها در خون مادر در اوایل بارداری، سرعت تمایز این سلول‌ها و نیمه‌عمر کوتاه آن‌ها در خون مادر و در نهایت، نشان‌گرهای اختصاصی‌ای که برای جدا سازی این سلول‌ها وجود دارد، اشاره نمود. به هر حال، میزان این سلول‌ها در خون مادر بسیار کم است که نیاز به تکنیک‌های مناسبی برای خالص سازی آن‌ها دارد. در حال حاضر، تکنیک‌هایی که به طور معمول مورد استفاده قرار می‌گیرند، شامل جداسازی سلولی به روش فعال‌سازی فلئورسنتی، جداسازی مغناطیسی و جداسازی بر اساس شیب بار می‌باشد. پس چنانچه سلول‌های جنینی جداسازی شوند، می‌توان از آن‌ها در کاربردهای بالینی همانند غربال‌گری برای ناهنجاری‌های کروموزومی به وسیله‌ی تکنیک هیبریداسیون فلئورسنتی درجا، یا تشخیص ناهنجاری‌های ژنی با استفاده از واکنش زنجیره‌ای پلیمریزاسیون استفاده نمود. 

کلیدواژه‌ها

عنوان مقاله [English]

Fetal Cells in Maternal Blood: Technical and Clinical Aspects

نویسندگان [English]

  • Meysam Mosallayi 1
  • Rasoul Salehi 2
1 MSc Student, Pediatric Inherited Diseases Research Center AND Department of Genetics and Molecular Biology, School of Medicine AND Students Research Committee, Isfahan University of Medical Sciences, Isfahan, Iran
2 Associate Professor, Pediatric Inherited Diseases Research Center AND Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
چکیده [English]

Currently, common used methods to obtain fetal material for prenatal diagnosis are potentially dangerous for fetus and mother; this has led to the effort by many groups to develop methods to recover fetal cells by non-invasive means, such as their enrichment from the peripheral blood of pregnant women. The fetal cell types studied by numerous investigators worldwide include fetal leukocytes, i.e. fetal lymphocytes and granulocytes, fetal nucleated red blood cells (NRBCs) and trophoblast cells. Fetal NRBCs have been the most commonly studied cell type, for this reason: the frequency of NRBCs in the fetus early in gestation is relatively high; these cells are also fairly well differentiated and likely to have a limited life span in the maternal circulation and finally, there are specific markers for the enrichment of these cells. However, the fetal cells in maternal blood are rare and a sophisticate technique is required for their enrichment; currently, the most commonly employed techniques are fluorescent activated cell sorting (FACS), magnetic cell sorting (MACS), and charge flow separation. Then, if the fetal cells can eventually be isolated, possible clinical applications will be included screening for fetal chromosome abnormalities via fluorescence in-situ hybridization (FISH) and for gene abnormalities by polymerase chain reaction (PCR).

کلیدواژه‌ها [English]

  • Fetal cells
  • Non-invasive
  • Maternal blood

مراجع

  1. Dey M, Sharma S, Aggarwal S. Prenatal screening methods for aneuploidies. N Am J Med Sci 2013; 5(3): 182-90.
  2. Kuliev AM, Modell B, Jackson L, Simpson JL, Brambati B, Rhoads G, et al. Risk evaluation of CVS. Prenat Diagn 1993; 13(3): 197-209.
  3. Purwosunu Y, Sekizawa A, Koide K, Okazaki S, Farina A, Okai T. Clinical potential for noninvasive prenatal diagnosis through detection of fetal cells in maternal blood. Taiwan J Obstet Gynecol 2006; 45(1): 10-20.
  4. Lapaire O, Holzgreve W, Oosterwijk JC, Brinkhaus R, Bianchi DW. Georg Schmorl on trophoblasts in the maternal circulation. Placenta 2007; 28(1): 1-5.
  5. Attwood HD, Park WW. Embolism to the lungs by trophoblast. J Obstet Gynaecol Br Commonw 1961; 68: 611-7.
  6. Hahn S, Holzgreve W. Fetal cells and cell-free fetal DNA in maternal blood: new insights into pre-eclampsia. Hum Reprod Update 2002; 8(6): 501-8.
  7. Sargent IL, Johansen M, Chua S, Redman CW. Clinical experience: isolating trophoblasts from maternal blood. Ann N Y Acad Sci 1994; 731: 154-61.
  8. Henderson KG, Shaw TE, Barrett IJ, Telenius AH, Wilson RD, Kalousek DK. Distribution of mosaicism in human placentae. Hum Genet 1996; 97(5): 650-4.
  9. Goldberg JD, Wohlferd MM. Incidence and outcome of chromosomal mosaicism found at the time of chorionic villus sampling. Am J Obstet Gynecol 1997; 176(6): 1349-52.
  10. Covone AE, Mutton D, Johnson PM, Adinolfi M. Trophoblast cells in peripheral blood from pregnant women. Lancet 1984; 2(8407): 841-3.
  11. Bertero MT, Camaschella C, Serra A, Bergui L, Caligaris-Cappio F. Circulating 'trophoblast' cells in pregnancy have maternal genetic markers. Prenat Diagn 1988; 8(8): 585-90.
  12. Hawes CS, Suskin HA, Kalionis B, Mueller UW, Casey G, Hall J, et al. Detection of paternally inherited mutations for beta-thalassemia in trophoblast isolated from peripheral maternal blood. Ann N Y Acad Sci 1994; 731: 181-5.
  13. Mueller UW, Hawes CS, Wright AE, Petropoulos A, DeBoni E, Firgaira FA, et al. Isolation of fetal trophoblast cells from peripheral blood of pregnant women. Lancet 1990; 336(8709): 197-200.
  14. Walknowska J, Conte FA, Grumbach MM. Practical and theoretical implications of fetal-maternal lymphocyte transfer. Lancet 1969; 1(7606): 1119-22.
  15. Schindler AM, Martin-du-Pan R. Prenatal diagnosis of fetal lymphocytes in the maternal blood. Obstet Gynecol 1972; 40(3): 340-6.
  16. Herzenberg LA, Bianchi DW, Schroder J, Cann HM, Iverson GM. Fetal cells in the blood of pregnant women: detection and enrichment by fluorescence-activated cell sorting. Proc Natl Acad Sci U S A 1979; 76(3): 1453-5.
  17. Tharapel AT, Jaswaney VL, Dockter ME, Wachtel SS, Chandler RW, Simpson JL, et al. Inability to detect fetal metaphases in flow-sorted lymphocyte cultures based on maternal-fetal HLA differences. Fetal Diagn Ther 1993; 8(2): 95-101.
  18. Bianchi DW, Zickwolf GK, Weil GJ, Sylvester S, DeMaria MA. Male fetal progenitor cells persist in maternal blood for as long as 27 years postpartum. Proc Natl Acad Sci USA 1996; 93(2): 705-8.
  19. Ciaranfi A, Curchod A, Odartchenko N. Post-partum survival of fetal lymphocytes in the maternal blood. Schweiz Med Wochenschr 1977; 107(5): 134-8. [In French].
  20. Clayton EM, Feldhaus WD, Whitacre FE. Fetal erythrocytes in the maternal circulation of pregnant women. Obstet Gynecol 1964; 23: 915-9.
  21. Holzgreve W, Garritsen HS, Ganshirt-Ahlert D. Fetal cells in the maternal circulation. J Reprod Med 1992; 37(5): 410-8.
  22. Bianchi DW, Flint AF, Pizzimenti MF, Knoll JH, Latt SA. Isolation of fetal DNA from nucleated erythrocytes in maternal blood. Proc Natl Acad Sci U S A 1990; 87(9): 3279-83.
  23. Miltenyi S, Muller W, Weichel W, Radbruch A. High gradient magnetic cell separation with MACS. Cytometry 1990; 11(2): 231-8.
  24. Price JO, Elias S, Wachtel SS, Klinger K, Dockter M, Tharapel A, et al. Prenatal diagnosis with fetal cells isolated from maternal blood by multiparameter flow cytometry. Am J Obstet Gynecol 1991; 165(6 Pt 1): 1731-7.
  25. Bianchi DW, Mahr A, Zickwolf GK, Houseal TW, Flint AF, Klinger KW. Detection of fetal cells with 47,XY,+21 karyotype in maternal peripheral blood. Hum Genet 1992; 90(4): 368-70.
  26. Ganshirt D, Garritsen H, Miny P, Holzgreve W. Fetal cells in maternal circulation throughout gestation. Lancet 1994; 343(8904): 1038-9.
  27. Troeger C, Zhong XY, Burgemeister R, Minderer S, Tercanli S, Holzgreve W, et al. Approximately half of the erythroblasts in maternal blood are of fetal origin. Mol Hum Reprod 1999; 5(12): 1162-5.
  28. Sohda S, Arinami T, Hamada H, Nakauchi H, Hamaguchi H, Kubo T. The proportion of fetal nucleated red blood cells in maternal blood: estimation by FACS analysis. Prenat Diagn 1997; 17(8): 743-52.
  29. Ganshirt-Ahlert D, Pohlschmidt M, Gal A, Miny P, Horst J, Holzgreve W. Ratio of fetal to maternal DNA is less than 1 in 5000 at different gestational ages in maternal blood. Clin Genet 1990; 38(1): 38-43.
  30. Hamada H, Arinami T, Kubo T, Hamaguchi H, Iwasaki H. Fetal nucleated cells in maternal peripheral blood: frequency and relationship to gestational age. Hum Genet 1993; 91(5): 427-32.
  31. Bianchi DW, Zickwolf GK, Yih MC, Flint AF, Geifman OH, Erikson MS, et al. Erythroid-specific antibodies enhance detection of fetal nucleated erythrocytes in maternal blood. Prenat Diagn 1993; 13(4): 293-300.
  32. Zheng YL, Demaria M, Zhen D, Vadnais TJ, Bianchi DW. Flow sorting of fetal erythroblasts using intracytoplasmic anti-fetal haemoglobin: preliminary observations on maternal samples. Prenat Diagn 1995; 15(10): 897-905.
  33. Simpson JL, Elias S. Isolating fetal cells in the maternal circulation. Hum Reprod Update 1995; 1(4): 409-18.
  34. Lewis DE, Schober W, Murrell S, Nguyen D, Scott J, Boinoff J, et al. Rare event selection of fetal nucleated erythrocytes in maternal blood by flow cytometry. Cytometry 1996; 23(3): 218-27.
  35. Bianchi DW, Klinger KW, Vadnais TJ, DeMaria MA, Shuber AP, Skoletsky J, et al. Development of a model system to compare cell separation methods for the isolation of fetal cells from maternal blood. Prenat Diagn 1996; 16(4): 289-98.
  36. Ganshirt-Ahlert D, Borjesson-Stoll R, Burschyk M, Dohr A, Garritsen HS, Helmer E, et al. Detection of fetal trisomies 21 and 18 from maternal blood using triple gradient and magnetic cell sorting. Am J Reprod Immunol 1993; 30(2-3): 194-201.
  37. Andrews K, Wienberg J, Ferguson-Smith MA, Rubinsztein DC. Enrichment of fetal nucleated cells from maternal blood: model test system using cord blood. Prenat Diagn 1995; 15(10): 913-9.
  38. Zheng YL, Carter NP, Price CM, Colman SM, Milton PJ, Hackett GA, et al. Prenatal diagnosis from maternal blood: simultaneous immunophenotyping and FISH of fetal nucleated erythrocytes isolated by negative magnetic cell sorting. J Med Genet 1993; 30(12): 1051-6.
  39. Wachtel SS, Sammons D, Manley M, Wachtel G, Twitty G, Utermohlen J, et al. Fetal cells in maternal blood: recovery by charge flow separation. Hum Genet 1996; 98(2): 162-6.
  40. Shulman LP, Phillips OP, Tolley E, Sammons D, Wachtel SS. Frequency of nucleated red blood cells in maternal blood during the different gestational ages. Hum Genet 1998; 103(6): 723-6.
  41. Hall JM, Adams S, Williams S, Rehse MA, Layton TJ, Molesh DA. Purification of fetal cells from maternal blood using an avidin-biotin immunoaffinity column. Ann N Y Acad Sci 1994; 731: 115-27.
  42. Steele CD, Wapner RJ, Smith JB, Haynes MK, Jackson LG. Prenatal diagnosis using fetal cells isolated from maternal peripheral blood: a review. Clin Obstet Gynecol 1996; 39(4): 801-13.
  43. Sitar G, Manenti L, Farina A, Lanati V, Mascheretti P, Forabosco A, et al. Characterization of the biophysical properties of human erythroblasts as a preliminary step to the isolation of fetal erythroblasts from maternal peripheral blood for non-invasive prenatal genetic investigation. Haematologica 1997; 82(1): 5-10.
  44. Ganshirt-Ahlert D, Burschyk M, Garritsen HS, Helmer L, Miny P, Horst J, et al. Magnetic cell sorting and the transferrin receptor as potential means of prenatal diagnosis from maternal blood. Am J Obstet Gynecol 1992; 166(5): 1350-5.
  45. Thilaganathan B, Meher-Homji NJ, Nicolaides KH. Blood transferrin receptor expression in chromosomally abnormal fetuses. Prenat Diagn 1995; 15(3): 282-4.
  46. Zheng YL, Zhen DK, Farina A, Berry SM, Wapner RJ, Williams JM, et al. Fetal cell identifiers: results of microscope slide-based immunocytochemical studies as a function of gestational age and abnormality. Am J Obstet Gynecol 1999; 180(5): 1234-9.
  47. Bianchi DW, Yih MC, Zickwolf GK, Flint AF. Transferrin receptor (CD71) expression on circulating mononuclear cells during pregnancy. Am J Obstet Gynecol 1994; 170(1 Pt 1): 202-6.
  48. Turpeinen U, Stenman UH. Determination of fetal hemoglobin by time-resolved immunofluorometric assay. Clin Chem 1992; 38(10): 2013-8.
  49. Von KH, Gahmberg N. Fetal erythroblasts from maternal blood identified with 2,3-bisphosphoglycerate (BPG) and in situ hybridization (ISH) using Y-specific probes. Prenat Diagn 1995; 15(2): 149-54.
  50. Hengstschlager M, Bernaschek G. Fetal cells in the peripheral blood of pregnant women express thymidine kinase: a new marker for detection. FEBS Lett 1997; 404(2-3): 299-302.
  51. Oosterwijk JC, Knepfle CF, Mesker WE, Vrolijk H, Sloos WC, Pattenier H, et al. Strategies for rare-event detection: an approach for automated fetal cell detection in maternal blood. Am J Hum Genet 1998; 63(6): 1783-92.
  52. Lo YM, Patel P, Wainscoat JS, Sampietro M, Gillmer MD, Fleming KA. Prenatal sex determination by DNA amplification from maternal peripheral blood. Lancet 1989; 2(8676): 1363-5.
  53. Lo YM, Patel P, Baigent CN, Gillmer MD, Chamberlain P, Travi M, et al. Prenatal sex determination from maternal peripheral blood using the polymerase chain reaction. Hum Genet 1993; 90(5): 483-8.
  54. Savion S, Carp H, Shepshelovich J, Irlin J, Kostykov M, Fein A, et al. Use of antibodies against the human antigen of erythroblasts for the detection of nucleated erythrocytes in the maternal circulation. Biol Neonate 1997; 71(2): 126-30.
  55. Troeger C, Holzgreve W, Hahn S. A comparison of different density gradients and antibodies for enrichment of fetal erythroblasts by MACS. Prenat Diagn 1999; 19(6): 521-6.
  56. Valerio D, Altieri V, Antonucci FR, Aiello R. Characterization of fetal haematopoietic progenitors circulating in maternal blood of seven aneuploid pregnancies. Prenat Diagn 1997; 17(12): 1159-69.
  57. Valerio D, Aiello R, Altieri V, Malato AP, Fortunato A, Canazio A. Culture of fetal erythroid progenitor cells from maternal blood for non-invasive prenatal genetic diagnosis. Prenat Diagn 1996; 16(12): 1073-82.
  58. Busch J, Huber P, Pfluger E, Miltenyi S, Holtz J, Radbruch A. Enrichment of fetal cells from maternal blood by high gradient magnetic cell sorting (double MACS) for PCR-based genetic analysis. Prenat Diagn 1994; 14(12): 1129-40.
  59. Ferguson-Smith MA, Zheng YL, Carter NP. Simultaneous immunophenotyping and FISH on fetal cells from maternal blood. Ann N Y Acad Sci 1994; 731: 73-9.
مجله دانشکده پزشکی اصفهان
دوره 32، شماره 313 - شماره پیاپی 313
بهمن و اسفند 1393
صفحه 2165-2173

فایل ها

سابقه مقاله

  • تاریخ دریافت: 13 خرداد 1393
  • تاریخ بازنگری: 11 آبان 1403
  • تاریخ پذیرش: 17 فروردین 1401

هم رسانی

ارجاع به این مقاله

آمار