Method for culturing liver stem cells

Abstract

The invention relates to a method for culturing liver stem cells. The method comprises the following steps of: a, flushing a liver tissue with PBS (Phosphate Buffered Saline) containing double antibodies for three times under the sterile condition; b, shearing the liver tissue, and digesting with a 0.25 percent g / mL trypsinase solution for 30 minutes; C, neutralizing digestive juice with a complete medium; d, beating and uniformly mixing the digested and neutralized liver tissue in a mixed liquid of the digestive juice and a neutralizing solution, centrifuging at the speed of 500 revolutions / minute for five minutes, extracting a supernatant liquid, and centrifuging the supernatant liquid at the speed of 1,500 revolutions / minute for five minutes; e, discarding the supernatant liquid, suspending precipitates with the complete medium, inoculating into a culture bottle, and culturing at the temperature of 37 DEG C in the environment of 5 percent CO2; f, culturing for four days, washing non-adherent cells, and changing the liquid with the complete medium; and g, changing the liquid with the complete medium once every four days, and performing passage in the ratio of 1:2 when the cells grow and expand to over 80 percent of the bottom area of the culture bottle.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and specifically relates to a method for extracting and expanding liver stem cells. Background technique [0002] In the past, the terminal treatment of liver cancer and advanced liver cirrhosis was liver transplantation, but the source of donors is scarce and there are many adverse reactions after transplantation. After the stem cell transplantation technology is applied in the clinic, it brings the hope of survival to the majority of patients. Initially, autologous bone marrow stem cells were used clinically, but the quality of bone marrow is affected by the patient's age, physical condition and other conditions, which often weakens the number and activity of stem cells. [0003] At present, the use of early (aborted) fetal tissue stem cells to treat related diseases has entered clinical practice. In 2003, the Ministry of Science and Technology and the Ministry of Health issued the "Guid...

AI Technical Summary

Problems solved by technology

This method takes a long time to process liver tissue and has many processing steps, which is easy to cause pollution and damage to cells

Differential centrifugation does not use digestive enzymes but cuts the tissue and removes the tissue fragments at different speeds to leave cells behind. However, this method contains more red blood cells, which will affect the attachment and growth of liver stem cells.

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