Human early placenta development model establishing method based on organs-on-chips

A technology of organ chips and developmental models, applied in biochemical equipment and methods, embryonic cells, microorganisms, etc., can solve problems such as the undetermined applicability of human hPSCs, and achieve great application value and strong repeatability

Active Publication Date: 2020-03-17
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although invaluable experience gained from the study of vertebrate development has helped us generate in vitro human embryos, the suitability of human hPSCs for modeling early development remains undetermined

Method used

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  • Human early placenta development model establishing method based on organs-on-chips
  • Human early placenta development model establishing method based on organs-on-chips
  • Human early placenta development model establishing method based on organs-on-chips

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Establishment of human early placental development model using hPSC self-assembled traits

[0026] The present invention uses a three-dimensional matrix material, which is a commercially available artificial basement membrane (Matrigel) (product number ref: 354277, company Corning).

[0027] A method for establishing a human early placenta development model based on an organ chip of the present invention adopts the above three-dimensional matrix material, and specifically includes the following process:

[0028] (1) Orifice plate modification and human pluripotent stem cell (hPSC) inoculation and culture

[0029] The 6-well plate was first modified with artificial basement membrane (Matrigel) and placed in a 4° refrigerator, then placed in a 37° incubator for 20 minutes, digested with cell digestion solution at room temperature, scraped off the cell mass with a cell scraper, and centrifuged at 600r for 2 minutes. The mTeSR1 medium containing Y27632 was resuspended to f...

Embodiment 2

[0037]Characterization of cell morphology and function differences between 2D and 3D matrices in an organ-on-a-chip-based model of human early placenta development

[0038] A human placenta development model was established using the same artificial basement membrane and cell seeding density as in Example 1. Here, the difference between hPSC induced differentiation under 2D conditions and 3D conditions in Case 1 is compared, which fully demonstrates that the model constructed in the three-dimensional matrix of the present invention has a nearly physiological three-dimensional structure, and is comparable to the traditional two-dimensional monolayer. In contrast, these cavities also showed efficient differentiation of trophoblasts.

[0039] The process of inducing hPSC differentiation under 2D conditions is as follows:

[0040] (1) Orifice plate modification and human pluripotent stem cell (hPSC) inoculation and culture

[0041] The 6-well plate was first modified with artifi...

Embodiment 3

[0048] In vivo establishment of an in vitro model of human early placental development using the self-assembly properties of hPSCs

[0049] The present invention uses a three-dimensional matrix material, which is a commercially available artificial basement membrane (Matrigel) (product number ref: 354277, company Corning).

[0050] A method for establishing an in vitro simulated human early placenta development model of the present invention adopts the above three-dimensional matrix material, and specifically includes the following process:

[0051] (1) Orifice plate modification and human pluripotent stem cell (hPSC) inoculation and culture

[0052] The 6-well plate was first modified with artificial basement membrane (Matrigel) and placed in a 4° refrigerator, then placed in a 37° incubator for 40 minutes, digested with cell digestion solution at room temperature, scraped off the cell mass with a cell scraper, and centrifuged at 600r for 1 minute. The mTeSR1 medium containi...

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Abstract

The invention discloses a human early placenta development model establishing method based on organs-on-chips, which mainly comprises the following steps of pore plate modification and human pluripotent stem cell (hPSC) inoculation culture, cell digestion and inoculation into a three-dimensional matrix, human early placenta development model establishment and the like. A near-physiological three-dimensional cavity structure is formed in a three-dimensional matrix material by utilizing the self-assembly property of hPSC, and the cavities also show the characteristic of efficient differentiationof trophoblasts. The method has the characteristics of simplicity, easiness in operation, high repeatability and the like. The establishment of the model can provide a platform for exploring human placentogenesis and placentogenesis in the field of cell level and molecular mechanism research, and also has great application value in epigenetics research in the placental development process.

Description

technical field [0001] The invention relates to the technical field of placental development, tissue bionics and cell biology research, in particular to a method for establishing a human early placental development model based on an organ chip. Background technique [0002] The placenta is an important organ that connects the mother and the fetus, and is also an important organ for material exchange between the fetus and the mother. The fetus develops in the womb and relies on the placenta to obtain nutrients from the mother to maintain a normal pregnancy. More and more studies have shown that diseases in human childhood and even adulthood are related to the growth and development of the placenta. Following the launch of the human placenta project in the United States, Chinese obstetrical experts have also paid more and more attention to the research on this key organ that conceives life. The research content involves many fields such as placental structure, infection and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073
CPCC12N5/0605C12N2513/00
Inventor 秦建华朱玉娟王慧
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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