Method for quantitatively evaluating proliferation of tumor cells transplanted into zebra fish embryos

Abstract

The invention belongs to the technical field of biology, and relates to a method for quantitatively evaluating proliferation of tumor cells transplanted into zebra fish embryos. The method is characterized by comprising the following steps of: performing fluorescence labeling on the tumor cells before injection; selecting the zebra fish embryos carrying the fluorescence tumor cells for further culture one day after injection; and amplifying human tumor cell genes by a quantitative PCR method 3 to 7 days after injection to evaluate the colonization and proliferation conditions of the human tumor cells in the zebra fish embryos. The injection period of the tumor cells subjected to fluorescence labeling in advance is the period of 48 hours after fertilization, and the injection part is a perivitelline space area. The injection volume of each embryo is 10 nL, and the number of the injected tumor cells is 300 to 500. During quantitative evaluation, a plurality of embryos are selected as a group generally, and the content of the human genes is detected by quantitative PCR. The period of 3 to 7 days after injection is an effective time window for evaluating whether the human tumor cells are proliferated in the embryos. The method is beneficial to the practicability of a PDX (patient-derived cell transplantation) model of zebra fish.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for quantitatively evaluating the proliferation of tumor cells transplanted into zebrafish embryos, which is characterized in that the tumor cells are fluorescently labeled before injection, and the zebrafish embryos carrying fluorescent tumor cells are selected one day after injection Further culture, and the human tumor cell colonization and proliferation in zebrafish embryos were evaluated by quantitative PCR method during the window period of 3 to 7 days after injection (equivalent to 5 to 9 days after fertilization). [0002] technical background [0003] It is generally believed that the occurrence of tumors is due to the cumulative effect of multiple gene mutations. Due to the diversity of gene mutations in cancer patients, existing tumor-targeted drugs can only be curative for cancer patients carrying a specific type of gene mutation. Therefore, for different tumor pat...

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Problems solved by technology

However, this method also has obvious disadvantages, that is, the fluorescent dyes will not increase with the proliferation of cells, and the fluorescent dyes on non-proliferated cell fragments still exist.

Therefore, direct observation of tumor cell proliferation cannot be achieved using this labeling method.

[0008] In addition, the allogeneic cell transplantation causes great damage to animal embryo cells. In practice, a considerable number of embryos die during the subsequent development of the embryo due to mechanical damage during cell transplantation, so they cannot be used as evaluation objects.

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