Villus tissue in-situ glass slide culture method and chromosome preparation method
An in situ culture and glass slide technology is applied in the field of villus tissue culture and detection, which can solve the problems of large cell loss, less complete chromosomal karyotype, and decreased karyotype analysis accuracy, achieves good dispersion, and improves diagnosis timeliness. Sexuality, the effect of simplifying the operation
Pending Publication Date: 2021-06-04
HUZHOU MATERNITY & CHILD CARE HOSPITAL
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Problems solved by technology
Compared with the direct method of villous cells, the culture bottle method has better quality and more quantity, but in the process of collecting cells, trypsinization method or scraping method is used, the loss of cells is large, the complete chromosome karyotype is less, and the accuracy of karyotype analysis is reduced.
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[0037] This embodiment provides a method for preparing chromosomes of villi tissue cells, comprising the following steps:
[0038] (1) In situ slide culture of villous tissue cells
[0039] ①Pre-treatment: Under sterile conditions, take 15 mg of villous branch samples as villous tissue samples, put them into a sterile petri dish, wash them with sterile saline until no visible blood red; use small tissue scissors and Check the villi with tweezers, remove other tissues, and ensure that the remaining villi are branched samples; add 5ml of double antibody solution to the sample, and then incubate in a carbon dioxide incubator at 37°C for 1 hour;
[0040] ② Digestion: Take out the petri dish, cut the villi tissue sample as finely as possible with small tissue scissors and tweezers, then transfer the villi to a sterile centrifuge tube with a sterile disposable pipette, centrifuge (1500rpm, 8 minutes), discard Supernatant; then add 4ml of fetal bovine serum medium (BI) to mix and wa...
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The invention belongs to the technical field of villus tissue culture and detection, and particularly discloses a villus tissue in-situ glass slide culture method and a chromosome preparation method. The villus tissue in-situ glass slide culture method comprises the steps of pretreatment, digestion, inoculation and culture, villus tissue cells are successfully cultured on a glass slide in situ, the glass slide can be directly used for preparing chromosomes after culture, compared with an existing culture bottle method, the cells do not need to be harvested through pancreatin digestion or scraping transfer, the operation is simplified, chromosome loss, cell loss and the like caused by experimental operation are avoided, and high-quality raw materials are provided for subsequent chromosome preparation. The chromosome preparation method is based on an in-situ glass slide culture method, cell loss is reduced to the minimum degree, the obtained chromosome is good in dispersity and complete in karyotype, and the karyotype analysis accuracy is effectively improved.
Description
technical field [0001] The invention belongs to the technical field of villous tissue culture and detection, and in particular relates to a villous tissue in situ glass slide culture method and a chromosome preparation method. Background technique [0002] According to statistics, the current incidence of birth defects in my country is about 4% to 6%, and nearly one million children with defects are born every year. In order to reduce the rate of birth defects, in the prenatal diagnosis of fetal diseases, amniocentesis, umbilical vein puncture and other methods are mainly used in the second and third trimesters. Due to the large gestational age of these methods, the serious abnormality of the fetus is found and the pregnancy is terminated later, the incidence of induced labor complications is higher, and it will cause great physical and psychological harm to the pregnant woman. With the development of medical technology, chorionic villus biopsy can now be performed in early...
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IPC IPC(8): G01N1/28G01N1/30
CPCG01N1/2813G01N1/30
Inventor 沈学萍阳鑫妙唐克峰
Owner HUZHOU MATERNITY & CHILD CARE HOSPITAL