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Stem cell preparation against malignant melanoma and preparation method thereof

A technology for malignant melanoma and stem cell preparations, which is applied in the field of biomedical research and can solve problems such as difficult to penetrate drug concentration, narrowing the treatment window, and clinical application limitations.

Active Publication Date: 2018-10-09
奥思达干细胞有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although traditional immunotoxins can also selectively target and kill tumor cells, due to their natural limitations: (A) it is difficult to penetrate into the tumor and reach a certain drug concentration; (B) the aggregation in non-target tissues is reduced The concentration of drug resistance is reduced and the therapeutic window is narrowed, so the clinical application is greatly limited

Method used

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  • Stem cell preparation against malignant melanoma and preparation method thereof
  • Stem cell preparation against malignant melanoma and preparation method thereof
  • Stem cell preparation against malignant melanoma and preparation method thereof

Examples

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specific Embodiment 1

[0040] 1. Preparation of competent cells (Escherichia coli DH5α, BJ5183) by calcium chloride method:

[0041] (1) Escherichia coli DH5α was streaked on the agar medium and cultured at 37°C for 16-20 hours;

[0042] (2) Pick a single colony about 5mm in size from the plate, inoculate it in 3-5ml LB medium, and culture it with strong shaking at 37°C and 200rmp for 12h until late logarithmic growth. Inoculate the bacterial solution in 100ml LB medium at a ratio of 1:50, and culture with vigorous shaking at 37°C and 200rpm for 3h until OD600=0.5;

[0043] (3) Place the culture on ice for 10 min, then transfer to two 50 ml centrifuge tubes, centrifuge at 4000 rpm, 4°C for 10 min;

[0044] (4) Discard the supernatant, invert the centrifuge tube for 1 min, remove the remaining liquid as much as possible, then add 10 ml of 0.05 mol / L CaCl pre-cooled with ice 2 Suspend the bacteria in the solution and place it on ice for 10 min;

[0045] (5) Centrifuge at 4000r / min for 10min at 4°C ...

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Abstract

The invention discloses a mesenchymal stem cell preparation capable of secreting specific recombinant immunotoxin Gas6-PE38KDEL for targeted therapy of malignant melanoma. A new therapy direction is provided for targeted therapy of the malignant melanoma. According to the mesenchymal stem cell preparation disclosed by the invention, a specific immunotoxin Gas6-PE38KDEL adenovirus vector composed of Axl ligand (Gas6) and a pseudomonas aeruginosa exotoxin derivative PE38KDEL is firstly constructed; a mesenchymal stem cell is infected in vitro; and the Gas6-PE38KDEL secreted by infected mesenchymal stem cell can be applied to specific targeted killing of malignant melanoma cells. The invention further provides a preparation method of the mesenchymal stem cell preparation.

Description

technical field [0001] The invention relates to a stem cell preparation against malignant melanoma and a preparation method thereof, belonging to the technical field of biomedical research. Background technique [0002] Malignant melanoma (MM) is a kind of skin tumor, and it is the fastest growing tumor among all malignant tumors, with an annual growth rate of about 3-5%. Malignant melanoma is formed by the malignant transformation of melanocytes located at the base of the epidermis, and mostly develops from moles or pigmented spots. Once it enters a rapid growth period, the prognosis is poor and the mortality rate is high, especially for metastatic malignant melanoma. The bit survival period is only about 5-11 months, the 1-year overall survival rate is only 27%, and the 5-year survival rate is less than 5%. 90% of malignant melanomas occur on the skin, most commonly on the back, chest and abdomen, and legs, and are not uncommon on the soles of the feet, fingers and toes, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775A61K35/28A61K35/12C12N15/861C12N15/31C12N15/12C12N5/10A61P35/00
CPCY02A50/30
Inventor 周萱
Owner 奥思达干细胞有限公司
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