Dielectrophoresis Activated Cell Sorting for Recovery of Fetal Erythroblasts

用于胎儿成红细胞回收的介电泳激活细胞分选

• 批准号: 7480545
• 负责人: Anna Asanbaeva
• 金额: $ 10.71万
• 依托单位: PHYSICAL OPTICS CORPORATION
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7480545
• 关键词: Address; Adult; Amniocentesis; Antibodies; Appearance; Biological Markers; Biotin; Birth; Blood; Blood Circulation; Buffers; Businesses; Cell Count; Cell Fractionation; Cell Separation; Cells; Cellular Morphology; Characteristics; Charge; Child health care; Chorionic Villi Sampling; Chromosomes; Clinical; Collection; Condition; Cultured Cells; DNA; Dependence; Detection; Development; Development Plans; Devices; Diabetes Mellitus; Diagnosis; Diagnostic Procedure; Down Syndrome; Early Diagnosis; Effectiveness; Electrodes; Erythroblasts; Erythrocytes; Evaluation; Fetus; Fluorescence; Fluorescent in Situ Hybridization; Frequencies; Funding; Future; Genetic; Goals; Government; Hematological Disease; Hospitals; Human Development; Immobilization; Immobilized Cells; In Situ Hybridization; Inborn Genetic Diseases; Institutes; Invasive; Kidney; Kidney Diseases; Lesion; Life; Magnetism; Mechanics; Medicine; Methods; Molecular; Molecular Abnormality; Molecular Analysis; Morphology; National Institute of Child Health and Human Development; Nature; Normal Cell; Numbers; Operative Surgical Procedures; Optics; Parents; Perinatal; Personal Satisfaction; Persons; Phase; Population; Pregnancy; Pregnant Women; Prenatal Diagnosis; Procedures; Process; Production; Public Health; Purpose; Rate; Recovery; Research; Rest; Risk; Sampling; Screening procedure; Sedimentation process; Simulate; Sorting - Cell Movement; Source; Spontaneous abortion; Standards of Weights and Measures; Support of Research; Surface; System; Techniques; Technology; Testing; Time; Tube; Umbilical Cord Blood; Uterus; Woman; base; commercialization; cost; density; design; design and construction; dielectric property; electric field; fetal; fetus cell; genetic analysis; improved; innovation; male; methylmalonic aciduria; miniaturize; neoplastic; novel; peripheral blood; prenatal; prototype; receptor; tumor; urologic

DESCRIPTION (provided by applicant): Current prenatal diagnostic methods are invasive in nature and carry significant risks of causing a miscarriage; therefore, they are not performed on all pregnant women. As a result, a large number of fetal genetic abnormalities, such as Down syndrome, are missed. Reliable methods of noninvasive prenatal diagnosis have long been sought in perinatal medicine. The presence of fetal nucleated red blood cells (NRBCs) in the maternal blood is well-established, and they are currently considered the best target for noninvasive prenatal screening and diagnosis. However, the detection of these cells remains problematic due to their low abundance in maternal circulation, and the presence of similar NRBCs that are of maternal origin. Current approaches for identification of fetal NRBCs rely on mechanical separation methods, such as flow-activated cell sorting (FACS) or magnetic-activated cell sorting (MACS), subsequent analysis of cellular morphology, and fluorescence in situ hybridization (FISH)- or PCR-based analyses of molecular or genetic biomarkers. However, according to the National Institute of Child Health and Human Development Fetal Cell Isolation Study (NIFTY) none of these methods have been shown to obtain fetal cells from maternal blood with sufficient reliability for routine prenatal diagnosis. Thus, the development of an efficient method for separating fetal NRBCs from maternal blood is necessary to enable the routine detection of a small numbers of these cells for noninvasive prenatal diagnosis. We hypothesize that the unique morphology of fetal NRBCs, and subsequently their unique dielectric properties, can facilitate separation of these cells by dielectrophoresis. This proposal describes an integrated design and development plan leading to a novel miniaturized Dielectrophoresis Activated Cell Sorting (DACS) system for recovery of fetal NRBCs from maternal peripheral blood. Isolation of fetal NRBCs with the DACS system will enable accurate, early diagnosis and screening for chromosomal and genetic abnormalities without risk of miscarriage. In addition, the DACS system can be adapted to separate other types of morphologically distinct cell populations from peripheral blood, such as those from a tumor or a pre-neoplastic lesion. PUBLIC HEALTH RELEVANCE: Successful development of the DACS technology will result in a highly sensitive and specific method for noninvasive identification and isolation of fetal NRBCs, allowing timely identification of chromosomal and genetic abnormalities in the fetus without the risks associated with standard procedures. Some of the identified prenatal conditions can then be treated to improve the outlook for the unborn baby. For example, biotin dependence and methylmalonic acidemia, both life-threatening inherited disorders, have been diagnosed by amniocentesis and treated in the womb, resulting in the births of healthy babies. When a fetus has a condition for which prenatal treatment is not yet possible, prenatal diagnosis permits parents to prepare themselves emotionally, and to plan the safest timing, hospital facility, and method of delivery.

描述（由申请人提供）：当前的产前诊断方法本质上是侵入性的，并具有引起流产的重大风险；因此，并非对所有孕妇进行。结果，错过了大量的胎儿遗传异常，例如唐氏综合症。长期以来一直在寻求非侵入性产前诊断的可靠方法。孕产妇血液中胎儿成核红细胞（NRBC）的存在是良好的，目前被认为是无创产前筛查和诊断的最佳目标。然而，由于这些细胞在母体循环中的丰度以及存在类似的母体起源的NRBC，因此对这些细胞的检测仍然存在问题。当前鉴定胎儿NRBC的方法依赖于机械分离方法，例如流动激活的细胞分选（FACS）或磁激活的细胞分选（MAC），随后对细胞形态学的分析以及原位杂交中的荧光（FISH） - 或基于PCR的分子或遗传生物标志物的分析。但是，根据国家儿童健康与人类发展研究所的胎儿细胞分离研究（Nifty）的说法，这些方法均未证明从孕妇血液中获得胎儿细胞，具有足够的可靠性来进行常规产前诊断。因此，必须开发一种有效的方法来将胎儿NRBC与母体血液分离，以便常规检测这些细胞以进行非侵入性产前诊断。我们假设胎儿​​NRBC的独特形态及其随后其独特的介电特性可以通过介电性型介电性促进这些细胞的分离。该提案描述了一项综合设计和开发计划，导致新型的微型介电介电细胞分选（DACS）系统，用于从母体外周血中恢复胎儿NRBC。用DACS系统分离胎儿NRBC将使染色体和遗传异常的准确，早期诊断和筛查，而不会流产风险。此外，DACS系统可以适应其他类型的形态学上不同类型的细胞群体与外周血（例如肿瘤或肿瘤前病变的血液）。公共卫生相关性：DACS技术的成功开发将导致一种高度敏感和具体的方法，用于胎儿NRBC的无创鉴定和隔离，从而及时鉴定胎儿的染色体和遗传异常，而没有与标准程序相关的风险。然后可以治疗一些确定的产前状况，以改善未出生婴儿的前景。例如，生物素依赖性和甲基化酸血症（两种威胁生命的遗传性疾病）已通过羊膜穿刺诊断并在子宫中进行治疗，从而导致健康婴儿的出生。当胎儿有可能无法治疗产前治疗的疾病时，产前诊断允许父母在情感上做好准备，并计划最安全的时机，医院设施和分娩方法。