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Slow-virus transfection for bone mesenchymal stem cells of machin

A technology of bone marrow mesenchyme and virus transfection, which is applied in the field of virus transfection efficiency of cells, can solve the problems of inability to study and reduce transfection efficiency, and achieve the effect of improving efficiency

Active Publication Date: 2013-04-24
BEIJING ZEPHYRM BIOTECHNOLOGY CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] However, in the genetic engineering experiment with the bone marrow MSCs of adult cynomolgus monkeys as the research object, the inventors found that adding antiretroviral drugs to the culture medium of the bone marrow MSCs of adult cynomolgus monkeys would make the cells undergo reverse transcription The transfection efficiency is greatly reduced in the process of viral transfection with virus as a vector, making it impossible to carry out related research

Method used

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  • Slow-virus transfection for bone mesenchymal stem cells of machin
  • Slow-virus transfection for bone mesenchymal stem cells of machin
  • Slow-virus transfection for bone mesenchymal stem cells of machin

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Embodiment 1 and comparative example 1 and 2

[0029] The inventor used lentivirus as a vector to transfect cynomolgus monkey mesenchymal stem cells. In Example 1, the cells used were adult cynomolgus monkey cells infected with simian foamy virus. , Stop using the antiretroviral drug Tenofovir on 6 and 8 days, that is, replace the medium 2, 4, 6 and 8 days before lentiviral transfection, and the replaced medium does not contain the antiretroviral drug Tenofovir Nofovir; in Comparative Example 1, the cells used were adult cynomolgus monkey cells infected with simian foamy virus, and the antiretroviral drug tenofovir was not stopped during the transfection process with lentivirus; In comparative example 2, the cells used were cynomolgus monkey cells not infected with simian foamy virus (these cynomolgus monkey cells were taken from young cynomolgus monkeys aged 1-3, and the experimental animal cynomolgus monkeys came from Lingkangsai, Nanning, Guangxi Nuoke Biotechnology Co., Ltd., the company's animal experiment facilities ...

Embodiment 1

[0049] The adult cynomolgus monkey bone marrow mesenchymal stem cells used in Example 1 and Comparative Example 1 were cryopreserved cells by the inventor, and were obtained from adult cynomolgus monkeys (these cynomolgus monkeys were from Lingkang Sanuoke Biotechnology Co., Ltd., Nanning, Guangxi). Co., Ltd., the company's animal experiment facilities have obtained the qualification certification of the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC), these cells have been used in the culture process before cryopreservation containing Thai Nofovir medium, if you use resuscitated adult cynomolgus bone marrow mesenchymal stem cells, stop using tenofovir after resuscitation, that is, use a medium without tenofovir, respectively, on the 2nd day after resuscitation. , 4, 6, and 8 days for viral transfection (referring to four parallel experiments for viral transfection at different time points, equivalent to discontinuing Tenofo on days 2, 4, 6, and ...

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Abstract

The invention relates to slow-virus transfection for bone mesenchymal stem cells of a machin, in particular to a method for improving virus transfection efficiency of cells infected by viruses. The method comprises the following steps of: 1) culturing the cells infected by the viruses; and 2) transfecting the cultured cells infected by the viruses. The method is characterized in that antiviral medicaments are added during the culturing of the cells infected by the viruses and are not used in a culture medium in a period of time before the transfection of the cultured cells. The method has theadvantage of greatly improving the virus transfection efficiency of the cells infected by the viruses based on an increased amplification amount.

Description

technical field [0001] The present invention relates to a method for improving the virus transfection efficiency of virus-infected cells, and more specifically, relates to a method for improving bone marrow mesenchymal stem cells of adult cynomolgus monkeys infected by Simian Foamy Virus (SFV). lentiviral transfection efficiency. Background technique [0002] Mesenchymal stem cells (MSCs) are mesoderm-derived stem cells with multi-directional differentiation capabilities, mainly found in connective tissues and interstitial organs, and most abundant in bone marrow tissues. differentiation potential of adult stem cells. Under certain induction conditions, MSCs have the ability to differentiate into mesoderm cells such as osteoblasts, chondrocytes, myoblasts, tenocytes, adipocytes and stromal cells; at the same time, MSCs can also differentiate into ectoderm neurons and endoderm hepatic oval cell differentiation. This breaks through the concept that MSCs are only mesoderm st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/867
Inventor 张愚任振华邹春林王淑艳岳峰
Owner BEIJING ZEPHYRM BIOTECHNOLOGY CO LTD