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Medical application of L-02 cells as cell source with liver function

A technology of L-02 and liver function, applied in the field of biotechnology and medicine, to achieve the effect of no risk of tumor formation and convenient source

Inactive Publication Date: 2012-04-04
TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether the L-02 cell line has liver cell-related functions and whether it can be applied to liver cell transplantation and bioartificial liver has not been reported in the domestic and foreign literature.

Method used

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  • Medical application of L-02 cells as cell source with liver function
  • Medical application of L-02 cells as cell source with liver function
  • Medical application of L-02 cells as cell source with liver function

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Liver cell function detection of embodiment 1L-02 cells

[0020] (1) Cell culture

[0021] Take out the previously frozen L-02 cells at -80°C, shake them in a water bath at 37°C for 1 minute to dissolve them, add them to a 15ml centrifuge tube, add a small amount of DMEM medium at the same time to remove the influence of DMSO, centrifuge at 1200rpm / min for 3 minutes, and remove Add 4ml of DMEM high-glucose medium, resuspend the cells, put them into a T25 cell culture flask, and place them in a 5% CO2 incubator at 37°C.

[0022] The medium was changed every 24 hours. After the cells grew to 80%-90% confluent in about 3 days, they were passaged at a ratio of 1:3. Subsequent experiments were performed after the cells expanded to a sufficient number.

[0023] Observing the cells under a microscope (X 100), it was found that the cells could adhere to the wall within 12 hours after thawing, and the cells were observed after 24 hours. The intercellular cells grew densely, wer...

Embodiment 2

[0125] In vivo application of embodiment 2L-02 cells

[0126] (1) Experimental grouping

[0127] G1 (n=10): intrasplenic injection of 5*10 7 L-02 cells, 90% liver resection 24 hours after liver cell transplantation;

[0128] G2 (n=15): Rats with 90% hepatectomy without hepatocyte transplantation.

[0129] (2) Intrasplenic transplantation of L-02 hepatocytes

[0130] S-D rats were anesthetized by intraperitoneal injection, fixed, and the abdominal skin was prepared. A midline abdominal incision was made to expose the spleen. Cell injections were performed directly on the parenchymal surface of the spleen. The experimental group will be 5×10 7 L-02 cells were suspended in 1ml DMEM medium and injected into the spleen, and the control group was injected with the same amount of DMEM medium. After the injection, press the spleen for several minutes to prevent bleeding and cell leakage, and then close the abdomen. After cell transplantation, FK506 was injected intramuscularly...

Embodiment 3

[0138] The safety verification of embodiment 3L-02 cell

[0139] Take 2×10 6 L-02 cells were implanted into the left shoulder of nude mice, and the same amount of Hepg-2 cells were implanted into the right shoulder of nude mice, and the tumor formation was observed after 4 weeks.

[0140] The results showed that nude mice had tumor formation on the right shoulder, but no tumor on the left shoulder (see Figure 8 ), indicating that L02 cells have better safety.

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Abstract

With the present invention, liver cell function research of a human fetal liver cell line L-02 is performed, and the human fetal liver cell line L-02 is adopted in liver cell transplantation. The research results show that: the L-02 cells have good liver cell functions, can express a plurality of liver cell specificity factors, wherein the liver cell specificity factors comprise albumin, bilirubin glucuronosyltransferase, blood coagulation factors, transaminase and the like. 90% hepatectomy-induced acute liver failure (ALF) rat model experiment results show that: the survival time of the ALF rats can be effectively prolonged with the L-02 cells, the tumorigenesis risk does not exist, the L-02 cells is expected to be the good cell source for liver cell transplantation, bioartificial liver, medical researches related to the liver cell functions, and the like, and is applicable for treatment of the ALF, liver cell function researches, and other medical fields.

Description

technical field [0001] The invention belongs to the field of biotechnology and medicine, and specifically relates to the study of liver cell function of L-02 cells, and its application in liver cell transplantation, treatment of acute liver failure, and medical research related to liver cell function after culture and proliferation in vitro . Background technique [0002] Acute liver failure (ALF) has a high mortality rate, and currently the only cure for acute liver failure is liver transplantation. However, the number of donated livers is far from meeting the needs of patients, and many patients with acute liver failure died while waiting for liver transplantation. In addition to liver transplantation, there are two effective treatments for acute liver failure, namely hepatocyte transplantation and bioartificial liver. Bioartificial liver filled with human hepatocytes is a recognized and effective method for the treatment of acute liver failure, can effectively reduce th...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61K35/407A61P1/16C12Q1/02
Inventor 周平胡昕鹏杨涛孟凡迎林菊生
Owner TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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