Prenatal screening

a technology for prenatal screening and fetal milk, applied in the field of prenatal screening, can solve problems such as unworkable approaches to da

Inactive Publication Date: 2006-01-26
BRANDEIS UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0013] One particular advantage of the present invention is its use in prenatal diagnosis. Activation of fetal cell nuclei can be used to facilitate prenatal diagnosis of various human conditions. Nuclei from of all types of human fetal cells including blood cells (such as red cells, white cells and other circulating cells of the fetus), as well as other types of fetal cells such as cells found in the amniotic fluid, or cells derived from the placenta (such as trophoblasts or syncytial trophoblasts), can be activated using the described products and methods. Preferably, the fetal cells to be activated are recovered from the blood or tissue of a pregnant woman rather than directly from the fetus or placenta, thereby decreasing the likelihood of discomfort or harm to the fetus and / or mother by the diagnosis procedure.

Problems solved by technology

After sporadic reports of success, recent articles appear to indicate that these markers are insufficiently specific and actually are attached to maternal cells frequently enough to make this approach unworkable to date.”

Method used

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Examples

Experimental program
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Effect test

example 1

Further Induction Optimization

[0235] This example describes additional experiments carried out to further determine the optimal induction time for an activating egg extract. Protein synthesis during the early part of the first cell cycle in activated eggs or egg extracts is required for preparation of an activating extract capable of efficient and complete genome replication. The required proteins can either be synthesized in intact eggs before preparation of extracts or in extracts including frozen / thawed extracts. As noted above, activating egg extract should be prepared from extracts induced for more than 10 minutes to enhance DNA synthesis activation activity.

[0236] It was found that proteins synthesized during the first 28-30 minutes in intact eggs (incubated at 20° C.) or during the first 60-80 minutes in a freshly prepared and activated extract (incubated at 25° C.), promote subsequent DNA replication. In contrast, proteins synthesized later in the first cell cycle, i.e., a...

example 2

cAMP Supplemented Activating Egg Extract

[0241] The affect of cAMP on DNA replication in activated Xenopus red blood cells was determined. Xenopus nuclei were isolated and pretreated by a method based on Coppock et al., Developmental Biology 131:102 (1989), as follows: Xenopus blood was obtained from females by cardiac puncture and collected using a syringe half-filled with Barth's solution (88 mM NaCl, 2.3 mM KCl, 0.82 mM MgCl2 and 10 mM Hepes, pH 7.4) containing heparin (10 mg / ml); the blood was immediately diluted into 10 ml of ice-cold 0.6×SSC (1×SSC is 0.15 M NaCl, 0.015 M Sodium citrate, pH 7.0) containing 0.1 mg / ml heparin, 0.1 mM TPCK (N-tosyl-L-phenylalanine chloromethyl ketone), 0.1 mM TLCK (Na-p-Tosyl-L-lysine chloromethyl ketone), 0.05 mM PMSF (phenylmethylsulfonyl fluoride), 5 μg / ml leupeptin, and 31.25 mM Na2S2O5; bleeds containing clots, even small ones, were rejected; diluted blood was underlaid with 0.5 volumes of ice cold Metrizamide® (refractive index of 1.3660 in...

example 3

Caffeine Supplemented Activating Egg Extract

[0245] The effect of caffeine on DNA replication in activated Xenopus red blood cells was determined. The experimental conditions used were as described in Example 1 with the following changes: the concentration of cAMP was set at 0.3 μM and caffeine was added to the activating egg extract to a concentration of either 0.2 mM, 1.0 mM, or 5.0 mM.

[0246] As illustrated by FIG. 1, caffeine at 1.0 mM in the presence of 0.3 μM cAMP gave the highest initial rate and extent of DNA replication in activated nuclei. Thus, caffeine can increase the activation activity of an activating egg extract.

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Abstract

The present invention concerns products and methods particularly useful for activating and analyzing non-dividing cell nuclei. The featured products include activating egg extracts, cytostatic factor (CSF) extracts, kits containing these extracts, and a microchamber microscope slide useful in analyzing nucleus activation.

Description

RELATED APPLICATION [0001] This application is a continuation-in-part of U.S. application Ser. No. 08 / 013,039 entitled “Prenatal Screening” the whole of which is hereby incorporated by reference herein (including the drawings).FIELD ON THE INVENTION [0002] This invention concerns products, methods and apparatus for analysis of non-dividing mammalian cell nuclei, such as human fetal cell nuclei and mammalian sperm cell nuclei. BACKGROUND OF THE INVENTION [0003] Jackson, Seminars in Perinatology 15:49 (1991), describes various procedures for prenatal diagnosis, including procedures to diagnose diseases. These procedures involve analysis of the DNA present in early embryonic stages. Specifically, Jackson mentions the use of a polymerase chain reaction to amplify genes, and the possibility of testing oocytes by polar body assay. According to Jackson: [0004]“There are other conceivable embryo biopsy approaches for prenatal diagnosis. The trophectoderm may be obtained at later, multicellu...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A01K31/19C12N15/85C12M3/04C12N5/02C12N5/076C12N15/873C12Q1/68C12Q1/6841C12Q1/70
CPCA01K2217/05C12N5/061C12N15/873C12N2500/80C12Q1/70C12N2501/70C12N2517/10C12Q1/6841C12N2501/01
Inventor WANGH, LAWRENCE J.
Owner BRANDEIS UNIV
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