Kit and method for cryopreserving induced pluripotent stem cells
A technology for pluripotent stem cells and cryopreservation, which is applied in the field of cryopreservation of induced pluripotent stem cells, can solve problems such as the need for improvement in cryopreservation methods, and achieve the effects of good cryopreservation effect, low pollution risk and good thawing effect.
Active Publication Date: 2018-08-03
HAINAN MEDICAL UNIV
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Problems solved by technology
[0003] However, current cryopreservation methods for cells, especially induced pluripotent stem cells, still need to be improved
Method used
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Embodiment 1
[0064] According to the method for cryopreserving induced pluripotent stem cells of the present invention, the experiment of cryopreserving human induced pluripotent stem cells was carried out with the open vitrification method as a control, as follows:
[0065] in,
[0066] iPSC: donated by Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences.
[0067] HFF (human foreskin fibroblast, human foreskin fibroblast): Provided by the Urology Department of the Affiliated Hospital of Hainan Medical College, isolated from the primary foreskin tissue, the foreskin was obtained from a circumcised child patient in the Urology Department of the Affiliated Hospital of Hainan Medical College , the specimens were used after the patients were informed and signed the informed consent.
[0068] 1 Experiment preparation
[0069] 1.1 Reagent preparation
[0070] 1.1.1 mTeSR TM 1 Preparation of working solution
[0071] iPSC medium mTeSR used in this experiment TM 1 (Ste...
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The invention discloses a kit and a method for cryopreserving induced pluripotent stem cells. The kit comprises: a first refrigeration working solution containing a first induced pluripotent stem cell medium, glycol and dimethyl sulfoxide (DMSO); and a second refrigeration working solution containing a second induced pluripotent stem cell medium, glycol, DMSO and sucrose, wherein the volume content of glycol in the first refrigeration working solution is same to the volume content of the DMSO in the first refrigeration working solution, the volume content of glycol in the second refrigeration working solution is same to the volume content of the DMSO in the second refrigeration working solution, and the volume content of glycol in the second refrigeration working solution is two times the volume content of the glycol in the first refrigeration working solution. The kit is safe and effective when the kit is used to cryopreserve the induced pluripotent stem cells (iPSCs), the thawing rate of the unfrozen iPSCs is high, and normal karyotype and characteristics can be maintained after culture passage.
Description
technical field [0001] The invention relates to a kit and a method for cryopreserving induced pluripotent stem cells. Background technique [0002] In 2006, Yamanaka et al applied reprogramming technology to induce mouse fibroblasts into pluripotent stem cells, also known as induced pluripotent stem cells (induced pluripotent stem cells, iPSCs). Compared with embryonic stem cells, iPSCs directly enter the state of pluripotent stem cells from somatic cells, avoiding ethical controversy; in addition, the discovery of iPSCs enables researchers to obtain stem cells from various cell sources of specific gene types in different patients, and apply them in clinical Treatment can reduce immune rejection. Therefore, iPSCs have attracted much attention in the fields of toxicology, pharmacology, tissue engineering and regenerative medicine. However, the widespread application of iPSCs in clinical practice still faces many problems, among which safety is the primary issue, that is, th...
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IPC IPC(8): A01N1/02
Inventor 马燕琳黄元华李崎蒋伟民张宇
Owner HAINAN MEDICAL UNIV