Preparation method of liver organoid

Abstract

The invention discloses a preparation method of liver organoid. The preparation method comprises the following steps: uniformly mixing acellular matrix hydrogel, stem cells and liver cells by using a serum-free culture medium; conducting stationary culture in a culture container until the acellular matrix hydrogel is solidified; and after it is determined that cells in the solidified acellular matrix hydrogel survive, continuing culture to obtain the liver organoid. The liver organoid prepared according to the method of the invention has ultralow immunogenicity and has the potential in cross-species transplantation.

Description

[0001] This application is a divisional application of the patent application "Method for Reprogramming Umbilical Cord Mesenchymal Stem Cells into Liver Cells and Prepared Liver Organoids". The filing date of the original application is August 28, 2020, and the application number is 2020108879718. The number is CN 112111444 A. technical field

[0002] The present application relates to a cell culture technology, in particular to a preparation method of liver organoids. Background technique

[0003] Liver tumors usually require major hepatectomy, and the remaining liver will regenerate to compensate, which can easily lead to posthepatectomy liver failure (PHLF), which is one of the most worrying complications and must be treated with transplantation and life-long immunization suppress to treat. The shortage of donor livers, high cost and immunosuppression limit the use of this treatment modality. Treatment of liver failure can also be improved with temporary liver support m...

Examples

Embodiment 1

[0048] Method for reprogramming umbilical cord mesenchymal stem cells into liver cells

[0049] Umbilical cord is derived from human beings, but it is usually treated as medical waste. Its sources are extensive and do not involve ethical issues. It is a superior tissue source for obtaining a large number of mesenchymal stem cells in clinical and research. Umbilical cord mesenchymal stem cells (UC-MSCs) have strong proliferative activity and have been used in the treatment of various diseases.

[0050] In this application, UC-MSCs were cultured to 6 points under serum-free conditions, that is, UC-MSCs were cultured in vitro until entering the logarithmic growth phase, such as figure 1 As shown, when UC-MSCs were cultured in vitro to the third passage, their morphology was uniform and they grew in the shape of a long spindle-shaped womb. Subsequent identification of cell surface markers and differentiation ability of UC-MSCs confirmed that the extracted UC-MSCs were in line with...

Embodiment 2

[0062] Liver organoid and preparation method thereof

[0063] Organoids are three-dimensional cell cultures that contain some of the key properties of their representative organs. Organoids are also an in vitro culture system that includes a self-renewing stem cell population that can differentiate into multiple organ-specific cell types, has a similar spatial organization to the corresponding organ and can reproduce some of the functions of the corresponding organ, thereby providing a Highly physiologically relevant system. Organoid cultures have been used in a variety of tissues, including gut, liver, pancreas, kidney, prostate, lung, optic cup, and brain. Organoids are also a tool with applications in developmental biology, disease pathology, cell biology, mechanisms of regeneration, precision medicine, and drug toxicity and efficacy testing, among others. For these applications and others, organoid culture provides a highly informative complement to existing two-dimensio...

[0001] This application is a divisional application of the patent application "Method for Reprogramming Umbilical Cord Mesenchymal Stem Cells into Liver Cells and Prepared Liver Organoids". The filing date of the original application is August 28, 2020, and the application number is 2020108879718. The number is CN 112111444 A. technical field

[0002] The present application relates to a cell culture technology, in particular to a preparation method of liver organoids. Background technique

[0003] Liver tumors usually require major hepatectomy, and the remaining liver will regenerate to compensate, which can easily lead to posthepatectomy liver failure (PHLF), which is one of the most worrying complications and must be treated with transplantation and life-long immunization suppress to treat. The shortage of donor livers, high cost and immunosuppression limit the use of this treatment modality. Treatment of liver failure can also be improved with temporary liver support m...

Examples

Embodiment 1

[0048] Method for reprogramming umbilical cord mesenchymal stem cells into liver cells

[0049] Umbilical cord is derived from human beings, but it is usually treated as medical waste. Its sources are extensive and do not involve ethical issues. It is a superior tissue source for obtaining a large number of mesenchymal stem cells in clinical and research. Umbilical cord mesenchymal stem cells (UC-MSCs) have strong proliferative activity and have been used in the treatment of various diseases.

[0050] In this application, UC-MSCs were cultured to 6 points under serum-free conditions, that is, UC-MSCs were cultured in vitro until entering the logarithmic growth phase, such as figure 1 As shown, when UC-MSCs were cultured in vitro to the third passage, their morphology was uniform and they grew in the shape of a long spindle-shaped womb. Subsequent identification of cell surface markers and differentiation ability of UC-MSCs confirmed that the extracted UC-MSCs were in line with...

Embodiment 2

[0062] Liver organoid and preparation method thereof

[0063] Organoids are three-dimensional cell cultures that contain some of the key properties of their representative organs. Organoids are also an in vitro culture system that includes a self-renewing stem cell population that can differentiate into multiple organ-specific cell types, has a similar spatial organization to the corresponding organ and can reproduce some of the functions of the corresponding organ, thereby providing a Highly physiologically relevant system. Organoid cultures have been used in a variety of tissues, including gut, liver, pancreas, kidney, prostate, lung, optic cup, and brain. Organoids are also a tool with applications in developmental biology, disease pathology, cell biology, mechanisms of regeneration, precision medicine, and drug toxicity and efficacy testing, among others. For these applications and others, organoid culture provides a highly informative complement to existing two-dimensio...

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