Construction method for hepatic progenitor cell-like cell bank and cell lines prepared by method and applications of cell bank and cell lines
A construction method and progenitor cell technology, applied in the construction method and its prepared cell lines and application fields, can solve the problems of affecting the in vitro proliferation ability of mature hepatic parenchymal cells, the single source of donors, and the inability to carry out drug-specific toxicity research, etc. Achieving good in vitro proliferative ability
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Embodiment 1
[0066] This example provides the first immortalized hepatic progenitor cell-like cell line, classified as 81.5, preserved in the China Center for Type Culture Collection, with the preservation number CCTCC NO: C2019125, and the first immortalized hepatic progenitor cell-like cell line. A method for constructing an immortal hepatic progenitor-like cell line.
[0067] The coating solution in this embodiment is a mixed solution of HepX medium and Matrigel, and the volume ratio of the HepX medium to the Matrigel is 80:1.
[0068] When the culture device is a 6-well cell culture plate, add 250-350 microliters of coating solution to each culture well to perform the coating treatment in advance.
[0069] When the culture device is a 12-well cell culture plate, add 150-200 microliters of coating solution to each culture well to perform the coating treatment in advance.
[0070] When the culture vessel is a cell culture dish with a diameter of 6 cm, add 500-700 microliters of coating ...
Embodiment 2
[0098] This embodiment provides a method for constructing a second immortal hepatic progenitor cell-like cell line and a second hepatic progenitor cell-like cell line obtained by the method for constructing a second immortal hepatic progenitor cell-like cell line.
[0099] The difference between the construction method of the second immortalized hepatic progenitor cell line and the construction method of the first immortalized hepatic progenitor cell-like cell line in Example 1 is that:
[0100] In the step S1, after separating and treating the human liver hemangiomatoma tissue derived from the second donor, the obtained second primary human hepatocyte culture was transformed and cultured for 14 days to obtain the second liver Progenitor-like cell lines. The inoculation density of the second human primary hepatocyte culture is 5×10 4 pieces / square centimeter. The number of times of expansion culture is 2, specifically the first expansion culture and the second expansion cult...
Embodiment 3
[0102] This embodiment provides a method for constructing a third immortal hepatic progenitor cell-like cell line and a third hepatic progenitor cell-like cell line obtained by the method for constructing a third immortal hepatic progenitor cell-like cell line.
[0103] The difference between the construction method of the third immortal hepatic progenitor cell-like cell line and the construction method of the first immortal hepatic progenitor cell-like cell line in Example 1 is that:
[0104] In the step S1, after the human liver hemangioma tissue from the third donor is separated and treated, the obtained third human primary hepatocyte culture is subjected to the transformation culture to obtain the third hepatic progenitor Cell-like cell line.
[0105] In the step S2, after thawing the frozen culture to be multiplied, 0.5×10 4 Inoculate at an inoculation density of one per square centimeter.
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