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A kind of preparation method of cryptorchid-specific germ cell model

A spermatogenic and specific technology, applied in the field of medical research, can solve the problem of limited research progress of research objects

Inactive Publication Date: 2015-12-16
SHANGHAI CHILDRENS HOSPITAL
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The limitations of research objects often limit the progress of research
Due to the constraints of ethics and experimental techniques, it is difficult to carry out the above researches on genes related to early development and reproduction in human body

Method used

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  • A kind of preparation method of cryptorchid-specific germ cell model
  • A kind of preparation method of cryptorchid-specific germ cell model
  • A kind of preparation method of cryptorchid-specific germ cell model

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Embodiment 1

[0028] Experimental program:

[0029] 1. Obtaining Seed Cells from Urine

[0030] 1) After being discussed and approved by the hospital ethics committee and the parents of the children signed the informed consent, 100-200ml of urine from children with cryptorchidism was collected for primary culture. After drinking water, collect 200-300ml of clean and interrupted urine, divide it into several 50ml centrifuge tubes in the ultra-clean bench, centrifuge at 1000rpm for 5min at room temperature, discard the supernatant, resuspend all the cells in the centrifuge tubes with 24ml of culture medium, and plant them on average in 24 wells In the plate, 1ml cell suspension / well, put it in the incubator, add fresh culture medium on the 3rd day, 1ml / well, replace the fresh culture medium on the 5th day, cells will appear after about 7-10 days, the cells It is short spindle-shaped, and grows in clusters at first. After the cells are fused to about 70-80%, they are continuously passaged, an...

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Abstract

The invention relates to the field of medical research, and relates to a preparation method of a cryptorchism specificity spermatogenic cell model. The preparation method of the cryptorchism specificity spermatogenic cell model, disclosed by the invention, comprises the following steps: collecting fresh urine of cryptorchism patients, and centrifuging and precipitating seed cells; adding seed cell culture fluid to the cells, and culturing; establishing and identifying iPS (induced pluripotent stem) cells; and inducing the iPS cells in vitro so as to differentiate the iPS cells into the spermatogenic cells. The preparation method has the advantages and characteristics that 1, the selected seed cells are cells derived from urine, and the sampling manner of the seed cells is completely non-invasive, easy to accept by children patients and very important for the research of children diseases; 2, various identifications prove that the iPS cells obtained from the urogenous cells have the characteristics similar to those of ES (embryonic stem) cells and can be induced in vitro and differentiated into the spermatogenic cells, and an experimental platform is supplied for the research of the molecular mechanism of cryptorchism morbidity and the drug screening.

Description

technical field [0001] The invention relates to the field of medical research, and relates to a method for preparing a cryptorchid-specific germ cell model. Background technique [0002] The testis is the main sex organ in males, and its function is to produce sperm, which combine with eggs to form new individuals. Cryptorchidism refers to the fact that the end of one or both testes descends to the scrotum and stays in any part of its normal descending path after birth. It is one of the most common congenital malformations of male genitalia, and its incidence has been on the rise in recent years. , and its molecular biological mechanism has not yet been clarified. One of the common complications of cryptorchidism is abnormal reproductive function in adulthood. If there is no early intervention for bilateral undescended testicles, most of them will cause male infertility in adulthood; but even if early intervention is carried out for children with cryptorchidism, such as bi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12N5/076
Inventor 陈方周君梅
Owner SHANGHAI CHILDRENS HOSPITAL