Isolating fetal trophoblasts

A trophoblast, fetal technology, applied in embryonic cells, tissue culture, cell dissociation methods, etc.

Inactive Publication Date: 2009-05-13
BIOCEPT INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although, the overall method described in this application basically provides an attractive method for prenatal diagnosis of underlying genetic diseases, there is still room for improvement in each step

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0056] The following base materials are preferably utilized:

[0057] Phosphate-buffered saline (PBS) containing BSA, pH7.4, (Sigma (Sigma) P-3688); 1M Tris-HCl, pH7.5, Celgro (Cellgro) (VWR45001-066); 1M magnesium chloride ( Sigma M-1028); Disodium hydrogen phosphate dihydrate (Sigma 71637); Sodium dihydrogen phosphate dihydrate (Sigma 71505); Pronase (50,000 U), (Calbiochem VWR 80601-406); β-galactosidase (1,500 U), (Roche 0 105 031); N-acetyl-L-cysteine, (Sigma A9165-25g); DNase I (150,000 U) (Sigma D-5033); sodium azide (Sigma S-8032); and Hams F-12 medium, HyClone (VWR 16777-488).

[0058] Prepare Specific Reagents

[0059] a. Phosphate buffer (0.2M phosphate / 1.5M NaCl, pH8.0) : Add 7.8 g sodium dihydrogen phosphate dihydrate, 8.9 g disodium hydrogen phosphate dihydrate, 43.83 g sodium chloride, and 450 ml sterile water into a 500 ml sterile bottle. The mixture was stirred with a magnetic stir bar until complete dissolution. Adjust the pH to 8.0 with 5M sodium hydro...

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Abstract

Methods for isolating and purifying fetal trophoblasts from a mucus sample obtained from the uterine cavity of a pregnant female. The mucus sample is transported from a clinical collection facility to a laboratory in a transportation medium so the cells remain viable. The mucus sample is then subjected to precise processing steps, including treatment with mucolytic agents or mucinases, sugar hydrolysis enzymes, nucleases, and proteases to provide fetal cells, the outer surfaces of which are so essentially completely devoid of attached mucosal biological material that they are then isolated in greater numbers than previously had been possible. The isolated cells are in appropriate condition to immediately be effectively subjected to FISH or to other molecular diagnostics.

Description

field of invention [0001] The present invention relates to methods of isolating fetal trophoblast (placental) cells obtained from pregnant female mammals, more particularly to the treatment of cervical mucus samples with trophoblast-releasing agents, and still more particularly to the Provides a method for fetal trophoblast samples that meet testing requirements such as FISH within approximately 8 hours of obtaining a sample from a pregnant female. Background of the invention [0002] Cells derived from a fetus can yield genetic and / or biochemical information about the fetus. By isolating trophoblast cells in the first trimester, these cells can be used to obtain fetal genetic and / or biochemical information, especially for detecting human fetal abnormalities. [0003] For many years, prenatal testing has been performed using fetal cells obtained by amniocentesis or chorionic villus sampling (CVS). Amniocentesis, usually performed at about 16 weeks of gestation, requires a ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N5/073
CPCC12N2509/00C12N5/0605
Inventor R·皮切尔R·法戈纳尼
Owner BIOCEPT INC
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