Culture apparatus special for in-situ culture of amniotic fluid/chorionic cells and application thereof

An in-situ culture and culture device technology, applied in the field of cell culture, can solve the problems of high technical requirements, great difficulty, and easy pollution, and achieve the effect of accurate chromosome variation, reasonable device design, and difficult judgment

Active Publication Date: 2013-03-13
浙江博圣生物技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because amniotic fluid cell culture has always had problems such as high technical requirements, great difficulty, long culture time, easy contamination, and few cleavage phases, many units still fail to carry out prenatal diagnosis of amniotic fluid
In the past 10 years, although the culture of amniocytes and the production of chromosomes have been continuously improved around the world, the culture and production of amniocytes are still difficult.

Method used

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  • Culture apparatus special for in-situ culture of amniotic fluid/chorionic cells and application thereof
  • Culture apparatus special for in-situ culture of amniotic fluid/chorionic cells and application thereof
  • Culture apparatus special for in-situ culture of amniotic fluid/chorionic cells and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] see figure 1 , the culture device is composed of a culture cover 1, a culture tank 2 and a glass slide 3, wherein a liquid filling hole 2a is set in the culture tank 2, which is convenient for adding liquid and changing the liquid, and will not affect the growth of the adherent cells. Two support bars 4a and 4b are arranged at the bottom of the culture tank 2, which not only play the role of earthquake resistance, but also facilitate access.

[0033] Wherein the glass slide 3 is surface-treated (siliconization treatment is to increase the adhesion of cells on the glass slide by forming a silicon-oxygen covalent bond on the glass slide, and the principle of collagen or polylysine treatment is to coat the surface of the glass slide with After being dyed with such substances, it is beneficial for amniotic fluid cells to adhere to the wall, preventing cells from swimming freely), and is suitable for cell adhesion and growth. The whole device is sterilized by cobalt 60 afte...

Embodiment 2

[0036] More than 2,000 cases of amniotic fluid / chorion specimens were used for cell culture and karyotype analysis with this culture device. A total of 1 fetus with Down syndrome (trisomy 21), 1 fetus with Patau syndrome (trisomy 13), and 1 fetus with Turner syndrome (X, 0) were detected. For the karyotype analysis results of Down syndrome, Patau syndrome, Turner syndrome and normal fetuses, see figure 2 , 3 , 4, 5.

[0037] Reagent material: the cell in situ culture dish described in the example;

[0038] Specimen: amniotic fluid / choriomembrane specimen from the antenatal clinic of Zhejiang Maternal and Child Health Hospital;

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Abstract

The invention provides a culture apparatus special for in-situ culture of amniotic fluid / chorionic cells, which consists of a culture tank, a slide and a culture cover. The culture tank is provided with a solution feeding hole to facilitate solution feeding and changing operation without influencing the growth of anchorage-dependent cells; and the bottom of the culture tank is provided with two supporting bars which play a role in resisting shock and are convenient to take and use. The amniotic fluid / chorionic cells are directly inoculated on the slide; a plurality of clones can be formed on one slide; and the subsequent analysis is also directly performed in situ of the clones. The culture apparatus effectively solves the problem that the cells are gathered in a concentrated way in the conventional culture bottle method and the conventional culture bottle method cannot identify which cloned cell population the chimera comes from. Because the cloned cells are separated from one another, the cells cloned with abnormal chromosomes and normal chromosomes can be counted; therefore, chromosomal variation of individual cells in isolated culture can be eliminated, the chimeric proportion can be accurately calculated, and the problem that the chimera is difficult to judge is solved.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and relates to an amniotic fluid / chorionic cell in situ culture dish, in particular to a culture device for providing a sterile environment for amniotic fluid / chorionic cell culture in prenatal diagnosis. Background technique [0002] Amniotic fluid / chorionic cell culture and chromosome preparation and analysis are the most commonly used methods for prenatal diagnosis of fetal congenital malformations and genetic diseases, and are the main means for prenatal diagnosis of chromosomal diseases. Most of the cells in the amniotic fluid are amniotic epithelium and fetal exfoliated epithelium, which come from fetal skin (including sweat glands and sebaceous glands), oral mucosa, part of the digestive tract, urethra, and reproductive tract, and endoderm epithelium from the larynx and trachea. After these cells are shed in the amniotic fluid, whether they can be cultured or grown is determined accor...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M3/04C12M1/22
CPCC12M23/38C12M47/04C12M23/04
Inventor 黄成刚王强张民
Owner 浙江博圣生物技术股份有限公司
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