Bionic liver organ culture model and application thereof

An organoid and model technology, which is applied in the field of in vitro cell culture technology and drug toxicity screening to achieve the effects of unique structure, enhanced toxicity sensitivity, and enhanced liver specific function and metabolic function.

Pending Publication Date: 2020-04-03

江苏信安佳医疗科技有限公司

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AI-Extracted Technical Summary

Problems solved by technology

However, there is currently no research on the co-culture model of hepatocytes and endothelial cells and its use in drug toxicity scre...

Abstract

The invention relates to the fields of cell in-vitro culture technology and drug toxicity screening, in particular to construction of a bionic liver organ culture model and an application of the bionic liver organ culture model in drug toxicity evaluation. The invention provides a construction method of the bionic liver organ culture model, which comprises the following steps: respectively carrying out conventional cell culture and digestion on hepatocytes and endothelial cells to prepare cell suspensions, respectively inoculating the cell suspensions to a groove device and a porous membrane,assembling the porous membrane to the groove device, and culturing in an incubator to prepare the bionic liver organ culture model. The model is unique in structure, has double cells and double targets, can be used for toxicity evaluation of hepatocytes with main targets and toxicity evaluation of non-parenchyma endothelial cells, and provides a new thought and a powerful tool for drug toxicity screening.

Application Domain

HepatocytesMicrobiological testing/measurement +3

Technology Topic

Organ culturePorous membrane +10

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Examples

Experimental program(4)
Effect test(1)

Example Embodiment

[0044] Example 1: Construction of a three-dimensional hepatocyte spheroid culture model alone (control group 1):

[0045] The hepatocytes were human hepatic progenitor cells HepaRG (purchased from Thermo Fisher), and the HepaRG cells were cultured according to the medium specified in the instructions (purchased from Thermo Fisher). HepaRG cells were digested with trypsin, and the cell viability was detected by trypan blue. The cell viability reached more than 90%, and it was ready for use.

[0046] Pipette the HepaRG cell suspension evenly and add it into the groove device, place it in a 37°C, 5% CO2 incubator for cultivation, and cultivate for 24 hours to form a three-dimensional hepatocyte spheroid culture model alone, and replace the culture every 2 days liquid.

Example Embodiment

[0047] Embodiment 2: Construction of endothelial cell culture model alone (control group 2):

[0048] The endothelial cells are human endothelial cells HUVEC (purchased from ATCC), and the HUVEC cells are cultured according to the medium specified in the instructions (purchased from ATCC). The HUVEC cells were digested with trypsin, and the cell viability was detected by trypan blue, and the cell viability reached more than 90%, and it was ready for use.

[0049] Pipette the HUVEC cell suspension evenly and add it to the tranwell, place it in a 37°C, 5% CO2 incubator for cultivation, and cultivate for 24 hours to form an endothelial cell culture model alone, and replace the culture medium every 2 days.

Example Embodiment

[0050] Example 3: Construction of a bionic liver organoid culture model (model group):

[0051] The hepatocytes were human hepatic progenitor cells HepaRG (purchased from Thermo Fisher), and the HepaRG cells were cultured according to the medium specified in the instructions (purchased from Thermo Fisher). The endothelial cells are human endothelial cells HUVEC (purchased from ATCC), and the HUVEC cells are cultured according to the medium specified in the instructions (purchased from ATCC). HepaRG and HUVEC cells were digested with trypsin, respectively, and the cell viability was detected by trypan blue. The viability of HepaRG and HUVEC cells reached more than 90%, and they were set aside.

[0052] Pipette the HepaRG cell suspension evenly and add it into the groove device, place it in a 37°C, 5% CO2 incubator for cultivation, and cultivate for 24 hours to form a three-dimensional hepatocyte spheroid culture model alone. The HUVEC cell suspension was pipetted evenly and added to the tranwell, placed in a 37°C, 5% CO2 incubator for cultivation, and cultured for 24 hours to form a single culture model of endothelial cells. Then, the groove device for culturing 3D hepatocyte spheroids and the transwell for culturing endothelial cells were reversibly assembled together to form a biomimetic liver organoid culture model. Mix HepaRG and HUVEC culture medium at a ratio of 1:1 to prepare the culture medium (model medium) for the bionic liver organoid culture model. The medium is prepared immediately before use, and the culture medium is replaced every 2 days.

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Description & Claims & Application Information

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Bionic liver organ culture model and application thereof

An organoid and model technology, which is applied in the field of in vitro cell culture technology and drug toxicity screening to achieve the effects of unique structure, enhanced toxicity sensitivity, and enhanced liver specific function and metabolic function.

AI-Extracted Technical Summary

Problems solved by technology

Abstract

Image

Examples

Example Embodiment

Example Embodiment

Example Embodiment

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Description & Claims & Application Information

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