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Novel in vitro model of human placental barrier

A human placenta and barrier body technology, applied to embryonic cells, vertebrate cells, animal cells, etc., can solve problems such as umbilical vessel stenosis, easily damaged tissues, and placental permeability that cannot be accurately evaluated by drugs

Inactive Publication Date: 2013-01-30
天津中医药大学第二附属医院
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Problems solved by technology

However, the limitations of this technique are that the umbilical vessels in the first and second trimesters are narrow and intubation is difficult, and the placental permeability of the drug cannot be accurately evaluated through this model. The operation of the placenta preparation and perfusion is complicated, and the tissue is easily damaged, which affects the results. the accuracy of
The primary culture and trophoblast cell lines (BeWo, Jar and JEG-3) of the placental trophoblast cell culture model have placental hormone synthesis and certain morphological characteristics of placental trophoblast cells, and can be inoculated on in vitro permeable supports However, this model cannot simulate the dynamic change process of the human placental barrier structure during pregnancy, and the experimental results obtained by cell models often need to be further confirmed by the placental perfusion method

Method used

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  • Novel in vitro model of human placental barrier

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Embodiment

[0033] 1. Preparation of reagents

[0034] 1. KRBB: Weigh NaCl 6.923g, KCl 0.354g, MgSO 4 ·7H 2 O 0.288g, CaCl 2 ·6H 2 O 0.56g, NaHCO 3 2.1g, Na 2 HPO 4 Mix 0.136g, glucose 1.8g, L-alanine 0.178g, fully dissolve with 800mL double distilled water, adjust the pH value to 7.4, make up the volume to 1000mL with double distilled water, pass through a 0.22μm filter membrane, aliquot, 4℃ save.

[0035] 2. Cryoprotectant solution A: Fully mix 7.5% ethylene glycol, 7.5% DMSO, and 85% pure water to prepare a solution.

[0036] 3. Cryoprotectant solution B: fully mix 15% ethylene glycol, 15% DMSO, and 70% pure water to make a solution.

[0037] 2. Establishment of human placental barrier in vitro model

[0038] 1. Preparation of Placental Tissue Thin Sections

[0039] 1) Collect the placenta: Select the placenta of healthy women who are 25-30 years old and have a full-term delivery at 36-40 weeks of gestation. After the placenta is delivered, wash the placenta three times wit...

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Abstract

The invention discloses a novel in vitro model of a human placental barrier, which stimulates an environment of a placenta in a parent body, and has a complete villus structure of the in vivo placenta, a secretion function and a material transport function; and the model still keeps the functional activity similar to the in vivo placenta after the model is continuously perfused for 5h. The model is built to provide a new rapid screening method for researching the structure, functions and material metabolism of the placenta and the relative diseases of the gestation, so that the human placental barrier has wide application value in the field of safety evaluation of medicines applied to toxicology, targeted therapy and the gestation period.

Description

Technical field: [0001] The invention relates to a novel human placenta barrier in vitro model. Background technique: [0002] Currently, there are two types of placental barrier in vitro models: placental perfusion model and trophoblast cell model. The placental perfusion model is to intubate the fetal arteries and veins in the complete placental lobules to establish fetal circulation, which can be used for the study of the placental transfer mechanism of exogenous and endogenous substances, and is used for placental metabolism and the transfer of endogenous substances between mother and fetus. Circulation, and a risk assessment of teratogenicity provide the most direct evidence. However, the limitations of this technique are that the umbilical vessels in the first and second trimesters are narrow and intubation is difficult, and the placental permeability of the drug cannot be accurately evaluated through this model. The operation of the placenta preparation and perfusion...

Claims

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Application Information

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IPC IPC(8): C12N5/073
Inventor 宋殿荣郭洁韩芳张崴王雅楠王玉华
Owner 天津中医药大学第二附属医院
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