Urine-derived mesenchymal stem cell mitochondria as well as transplanting method and application thereof

A technology of stem cells and mitochondria, applied in the field of biomedicine, to achieve the effect of unlimited sources, solving the problem of heterogeneity and ethics, and improving quality

Pending Publication Date: 2022-03-04
PEOPLES HOSPITAL PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on the use of urine-deriv

Method used

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  • Urine-derived mesenchymal stem cell mitochondria as well as transplanting method and application thereof
  • Urine-derived mesenchymal stem cell mitochondria as well as transplanting method and application thereof
  • Urine-derived mesenchymal stem cell mitochondria as well as transplanting method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0057] This embodiment mesenchymal stem cells obtained between the urine source mitochondria extracted by the following method:

[0058] (1) to a urine collection vessel, centrifuged, the supernatant discarded, PBS buffer was added to the vessel were resuspended, centrifuged again between the supernatant was discarded, urine-derived mesenchymal stem cells isolated cell pellet was resuspended culture medium, seeded gelatin-coated six-well plates, placed in primary culture incubator; after the whole was changed once a small colony formation, to be cloned after integration into a large, using trypsin digestion, trypsin digestion aspirated, amplification using USC resuspended medium, inoculated into the new six-well plates, referred to as P1 generation (hereinafter passaged and cultured and the same);

[0059] Obtained when (2) the step of (. 1) is placed in an incubator on behalf of P1 culture was continued until the six-well plates P1 generations covered 95% of the area, using tryps...

Embodiment 2

[0062] This embodiment provides a mesenchymal stem cell mitochondria inter urinary allograft method comprising: in intracytoplasmic sperm injection period, the mesenchymal stem cells and the sperm mitochondria between the source of urine (using mitochondrial preservation solution mitochondrial pellet was resuspended the resulting solution droplets mitochondria) co-injected into the mature oocytes, blastocysts cultured performed to improve the quality of oocytes, fertilization rate and embryo quality increase, improve human embryo development in vitro fertilization rate.

Embodiment 3

[0064] Mitochondrial bioactive mature oocytes low prognosis of patients in the present embodiment is compared with the normal population

[0065] Confocal microscope mitochondrial activity of the mature oocytes with low normal population prognosis, the TMRE to tetramethylrhodamine ethyl ester, i.e., an indicator of mitochondrial membrane potential, mitochondrial activity representative of the intensity of red fluorescence and the results are figure 1 Indicated.

[0066] from figure 1 The results showed that normal biological activity of the mitochondrial population oocytes was significantly higher than patients with low prognosis.

[0067] By the following steps (1) - between the urine extracted source (3) to give the same low prognosis mesenchymal stem cells mitochondria:

[0068] (1) the prognosis of patients with low urine collected above 200 mL, 50mL dispensed into sterile centrifuge tubes, 1200 rpm for 10 minutes, the supernatant was discarded, 20 ml of PBS and resuspended ag...

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Abstract

The invention relates to urine-derived mesenchymal stem cell mitochondria and a transplantation method and application thereof.The urine-derived mesenchymal stem cell mitochondria is extracted from urine and used for improving the quality of oocytes, and the transplantation method comprises the steps that in the single sperm microinjection period in oocytoplasm, the single sperm is inoculated into the oocytoplasm; the sperms and mitochondria of the urine-derived mesenchymal stem cells are jointly injected into mature oocytes, and blastocyst culture is carried out. The method has the beneficial effects that the fertilization rate and embryo quality of human in-vitro fertilization are obviously improved by combining the transplantation of the mitochondria of the urine-derived mesenchymal stem cells in the traditional ICSI period, and the method can be used for in-vitro fertilization of infertility patients with low prognosis and has a better treatment effect. The problem of source of autologous mitochondria in the prior art is solved, and introduction of third-party genetic materials and ethics are not involved; the transplantation method is easy to operate, high in feasibility and capable of being used for autologous noninvasive treatment.

Description

Technical field [0001] The present invention belongs to the field of biomedical technology, involving a mitochondria between urine gamma mesenchymal stem cell mitochondria and its transplantation method, specifically, interposed therein, a synthetic source of autologous non-invasive sources during micro-injection (ICSI). Meikid of the mitochondrial mitochondrial mitochondrial transplantation method can be used to improve the quality of oocytes and the cultivation of in vitro and embryo. Background technique [0002] In the large environment of the country's fertility and continuous decline and the continuous reproduction of birth age, it is a research hot spot in the reproductive field. In vitro fertilization / single-spermum erytoplasm injection technology (IVF / ICSI) is the effective measures currently in the treatment of infertility, and international success rate is about 40%, but for elderly Patients or people with poor egg / embryonic quality, that is, low-resembling crowd...

Claims

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Application Information

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IPC IPC(8): C12N5/0775C12N15/89C12N5/073C12N5/075
CPCC12N5/0668C12N15/89C12N5/0604C12N5/0609C12N2509/00C12N2533/54A61P15/08C12N5/0608C12N2506/1392C12N5/061C12N2517/10
Inventor 沈浣姜之歆石程王艳槟陈曦韩红敬付闵费嘉
Owner PEOPLES HOSPITAL PEKING UNIV
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