Method for reprogramming umbilical cord mesenchymal stem cells into liver cells and prepared liver organoids

A technology of liver cells and stem cells, applied in the field of liver organoids, can solve problems such as little effect, and achieve the effects of increased albumin expression, easy preparation, and improved induction efficiency

Active Publication Date: 2020-12-22
GUANGDONG UNISUN BIOTECHNOLOGY CO LTD
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, several traditional approaches used to date have had little success, and these hepatocyte-like cells exhibit only a subset of the markers and functions of primary hepatocytes.

Method used

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  • Method for reprogramming umbilical cord mesenchymal stem cells into liver cells and prepared liver organoids
  • Method for reprogramming umbilical cord mesenchymal stem cells into liver cells and prepared liver organoids
  • Method for reprogramming umbilical cord mesenchymal stem cells into liver cells and prepared liver organoids

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Embodiment 1

[0047] Method for reprogramming umbilical cord mesenchymal stem cells into liver cells

[0048] 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.

[0049] 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

[0061] Liver organoid and preparation method thereof

[0062] 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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Abstract

The invention discloses a method for reprogramming umbilical cord mesenchymal stem cells into liver cells. The method comprises the following steps of obtaining umbilical cord mesenchymal stem cells;carrying out pretreatment on the umbilical cord mesenchymal stem cells for two days; carrying out liver formation induction on the pretreated umbilical cord mesenchymal stem cells for seven days by using a liver formation induction composition; carrying out primary maturation induction on the umbilical cord mesenchymal stem cells subjected to liver formation induction treatment by using a first maturation induction composition for seven days to obtain liver cells; and carrying out second maturation induction on the liver cells for 12-20 days by using a second maturation induction composition to obtain the functional liver cells. Liver organoids comprise acellular matrix hydrogel, stem cells and liver cells, wherein the stem cells and liver cells being distributed in the acellular matrix hydrogel. According to the reprogramming method, the step of secondary maturation induction is added, so tat the culture time is prolonged, and the induction efficiency is remarkably improved. The liverorganoids provided by the invention have ultra-low immunogenicity and the possibility of cross-species transplantation.

Description

technical field [0001] The present application relates to a cell culture technology, in particular to a method for reprogramming umbilical cord mesenchymal stem cells into liver cells and the prepared liver organoids. Background technique [0002] 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 methods. For example, bioartificial liver, continuous hemodiafiltration through plasma exchange and albumin dialysis can improve some liver function parameters, but it is difficult to prove that it can improve patient survival. And the technique has been l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/0775
CPCC12N5/067C12N2501/11C12N2501/115C12N2501/12C12N2500/90C12N2533/90C12N2506/1392C12N2500/30C12N2501/999C12N2501/30
Inventor 高毅易笑陈枫李阳王沂峰刘凡郜坤洁
Owner GUANGDONG UNISUN BIOTECHNOLOGY CO LTD
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