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Cell therapeutic agent for cancer treatment and combination therapy with same

A cancer treatment and cell technology, applied in the direction of genetically modified cells, cell culture active agents, animal cells, etc., can solve the problem of not doing too much research, and achieve the effect of maximizing the effect and excellent tumor inhibition effect.

Pending Publication Date: 2017-12-08
CELL & BRAIN CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in combination therapy using mesenchymal stem cells, studies aimed at administering cells such as autologous stem cells to restore cell damage after anticancer treatment by chemotherapeutic agents have been intensively conducted, but there is little interest in using stem cells themselves as combination therapy Therapeutics for have not been studied much

Method used

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  • Cell therapeutic agent for cancer treatment and combination therapy with same
  • Cell therapeutic agent for cancer treatment and combination therapy with same
  • Cell therapeutic agent for cancer treatment and combination therapy with same

Examples

Experimental program
Comparison scheme
Effect test

preparation Embodiment 1

[0073] Preparation Example 1: Preparation of Mesenchymal Stem Cells Expressing Cytosine Deaminase (CD)

[0074] 1.1 Isolation and culture of mesenchymal stem cells

[0075]4 ml of human bone marrow donated after IRB examination at Ajou University Medical Center was overlaid with 4 ml of HISTOPAQUE 1077 (Sigma-Aldrich) in a sterile 15 ml tube and centrifuged at 400 x g for 30 at room temperature. minute. After centrifugation, 0.5 ml of the intermediate buffy coat was carefully collected using a Pasteur pipette and, after centrifugation at 250 x g for 10 minutes, transferred to a test tube containing 10 ml of sterile phosphate buffered saline (pH 7.4) , discard the supernatant, and add 10 ml of phosphate buffered saline, and gently suspend, then centrifuge at 250 x g for 10 min. This operation was repeated twice, and the final pellet was added to DMEM medium (Gibco) supplemented with 10% FBS (like standard fetal bovine serum (Hyclone)) and dispersed in 100 mm animal cell cultu...

Embodiment 1

[0082] Suicide effect and bystander effect of embodiment 1.MSC / CD

[0083] 1.1 Confirm the suicide effect

[0084] Cytosine deaminase (CD) is a suicide gene. MSC / CD, which are mesenchymal stem cells expressing cytosine deaminase (CD), can convert the prodrug 5-fluorocytosine (5-FC) to 5-fluorouracil (5-FU), and can have figure 1 Suicide effect (cells killing themselves) and bystander effect (killing of surrounding cells) are shown. Therefore, the suicide effect and bystander effect of MSC / CD prepared in specification 1.3 were eliminated. First, 10,000 MSC / CD or mesenchymal stem cells (MSC) were cultured in a 12-well plate. From the next day, cells were treated with prodrug 5-FC at a concentration of 0-1,000 μM, and replaced every two days with new medium containing the drug. Change medium to cell culture medium containing 0.5 mg / ml MTT (3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide, Sigma-Aldrich) A base capable of measuring live cells on day 6 after treat...

preparation Embodiment 2

[0090] Preparation Example 2. Preparation of "I cell" and "F cell"

[0091] In the case of cell therapeutics, the effect can vary under various conditions according to changes in the state of the cells. Therefore, in order to confirm whether cell ability changes depending on the state of MSC / CD, cells in a state of thawing immediately after freezing (hereinafter, referred to as F cells (frozen cells)), and cells in a state of collecting from cells immediately during culture (hereinafter, referred to as F cells) were prepared. for I cells (cells collected immediately)).

[0092] To prepare F cells and I cells separately, MSC / CD was cultured, and then 2×10 6 Disperse each cell in 1ml freezing medium containing 2%-10% (2%, 5%, 10%) DMSO or 2×10 6 The cells were dispersed in 1 ml of CryostorCS10 (BioLife Solutions). Cells were frozen and stored in liquid nitrogen containers. First, to obtain "I cells" of MSC / CD, cells from passage 5 were lysed at 37°C and placed in 9 ml of cul...

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Abstract

The present invention relates to a method for preparing cells for cancer treatment and a kit for cancer treatment comprising cells prepared by the method. The preparation method of the present invention can provide F cells, which, in spite of having no difference in the proliferative capacity compared with mesenchymal stem cells expressing cytosine deaminase, which are harvested and used immediately after the culture, exhibit a very excellent tumor suppressive effect through the treatment together with 5-FC and induce a remarkable synergistic effect exceeding an effect from combinative treatment with an existing anticancer drug in cases of a combinative treatment with another anticancer drug. Therefore, the present invention can be utilized for a kit for cancer treatment comprising such F cells, and thus can be favorably used to maximize the effect of existing cancer treatments.

Description

technical field [0001] The present disclosure relates to a method for preparing cells for cancer treatment, an anticancer adjuvant comprising the cells prepared by the method, a kit for cancer treatment, and a method for cancer treatment. Background technique [0002] Millions of people around the world die from various types of cancer, including: bone cancer, bladder cancer, blood cancer (leukemia), brain cancer, breast cancer, colorectal cancer, cervical cancer, esophagus cancer, bowel cancer, kidney cancer , lung cancer, liver cancer, oral cancer, nasal cancer, nerve cancer, ovarian cancer, pancreatic cancer, skin cancer, stomach cancer, prostate cancer, neck cancer, uterine cancer and vaginal cancer. Over the years, several methods including radiation and chemotherapy have been used to treat cancer, but the number of cancer patients is still increasing. It has been widely reported that radiation and chemotherapy can lead to toxicity-related diseases and even death in so...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/12A61K31/513A61K45/06A61P35/00A61K33/243A61K33/244
CPCA61K31/513A61K35/12A61K45/06A61K33/243A61K33/244A61K2300/00A61K31/4188A61K31/4745A61K31/506A61K31/675A61K31/704C12N5/0663C12N2510/00C12N2523/00A61P1/00A61P1/02A61P1/04A61P1/16A61P1/18A61P11/00A61P11/02A61P13/08A61P13/10A61P13/12A61P15/00A61P17/00A61P25/00A61P35/00A61P35/02A61P43/00A61K35/28C12N5/0665C12N2501/73
Inventor 徐海荣金性洙张多荣郑镇花李永敦黄宇燮朴真成李秀贞
Owner CELL & BRAIN CO LTD
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