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Method for inducing tumor cells to form tumor stem cells

A tumor stem cell and tumor cell technology, applied in the field of inducing tumor cell lines to form tumor stem cells, to achieve the effects of improving adaptability, breaking the limitation of consumables, and reducing adhesion ability

Pending Publication Date: 2022-05-13
WUHAN PROCELL LIFE SCI & TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Based on this, the present invention provides a new method for inducing tumor cells to form tumor stem cells, so as to solve the problems in the existing sphere-forming culture mentioned in the background art

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  • Method for inducing tumor cells to form tumor stem cells

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Embodiment 1

[0034] This embodiment provides a method for inducing tumor cells to form tumor stem cells, specifically using HepG2 (human liver cancer cells) as experimental cells, including the following steps:

[0035] (1) Prepare tumor stem cell induction medium: add 2% B27 cell supplement and 1% P / S (penicillin-streptomycin mixture) to DMEM / F12 medium at a concentration of 20 ng / mL RecombinantHuman LIF (human leukemia inhibitory factor), 20ng / mL of Recombinant Human FGFb (human basic fibroblast growth factor) and 20ng / mL of Recombinant Human EGF (human epidermal growth factor).

[0036] (2) Take tumor cells in good growth state in the logarithmic growth phase.

[0037] (3) Inoculation: After preparing the experimental materials, the tumor cells in the logarithmic growth phase were inoculated into the cell culture dish.

[0038] (4) Serum-free synchronization: After the cells enter the logarithmic growth phase, add serum-free DMEM / F12 medium and perform synchronization for 12 hours.

...

Embodiment 2

[0044] This embodiment provides a method for inducing tumor cells to form tumor stem cells, specifically using HCCLM3 (human highly metastatic liver cancer cells) as experimental cells, including the following steps:

[0045] (1) Prepare tumor stem cell induction medium: add 2% B27 cell supplement and 1% P / S (penicillin-streptomycin mixture) to DMEM / F12 medium at a concentration of 20 ng / mL RecombinantHuman LIF (human leukemia inhibitory factor), 20ng / mL of Recombinant Human FGFb (human basic fibroblast growth factor) and 20ng / mL of Recombinant Human EGF (human epidermal growth factor).

[0046] (2) Take tumor cells in good growth state in the logarithmic growth phase.

[0047] (3) Inoculation: After preparing the experimental materials, the tumor cells in the logarithmic growth phase were inoculated into the cell culture dish.

[0048] (4) Serum-free synchronization: After the cells enter the logarithmic growth phase, add serum-free DMEM / F12 medium and perform synchronizatio...

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Abstract

The invention particularly relates to a method for inducing tumor cells to form tumor stem cells. The specific method comprises the following steps: performing synchronization treatment on tumor cells in a logarithmic phase, inoculating the tumor cells in a suspension state into a tumor stem cell induction culture medium, periodically applying mechanical force at fixed time intervals to suspend, dispersing the adherent tumor cells, and continuing to culture. According to the method disclosed by the invention, the adhesion capacity of the cells is reduced through the methods of synchronous serum-free treatment, suspension state inoculation and periodic suspension dispersion, so that the cells directly enter a suspension state, and the screening process of the dry tumor cells is completed only by using a common culture dish, a pipette, a centrifugal machine and the like; high-quality experiment results are achieved, meanwhile, limitations of valuable consumables, advanced equipment and complex technologies are broken through, and experiment cost, equipment limitations and technical thresholds are reduced.

Description

technical field [0001] The invention relates to the technical fields of tumor medicine and cell biology, in particular to a method for inducing tumor cell lines to form tumor stem cells. Background technique [0002] Studies in recent years have confirmed that there are a small number of stem cell-like cell populations in tumor tissues. These cells have self-renewal ability and multi-directional differentiation potential, which are called tumor stem cells; tumor stem cells may be the root cause of malignant tumor recurrence and metastasis . The vast majority of solid tumors are considered to originate from tumor stem cells, which may be one of the important causes of primary and acquired drug resistance in clinical treatment of tumors. extensive research attention. However, there are still many factors restricting the research of tumor stem cells. One of the key factors is that the cultivation and screening technology of tumor stem cells is not mature enough to conduct ext...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/095
CPCC12N5/0695
Inventor 冷毅斌吴娟吴亭李韦甫
Owner WUHAN PROCELL LIFE SCI & TECH CO LTD
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