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Establishment method of urine cell induced pluripotent stem cells of acute kidney injury patient

A technology of acute kidney injury and pluripotent stem cells, applied in the field of establishment of induced pluripotent stem cells, which can solve the problems of unobtainable and low efficiency of kidney cells

Active Publication Date: 2021-12-03
NANJING MEDICAL UNIV
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Problems solved by technology

At present, urine kidney cells are mainly derived from newborns, premature infants and fetuses with immature kidneys, while the efficiency of obtaining kidney cells from the urine of children and adults with mature kidneys is extremely low or even impossible.
Although improved efficiency of cell isolation from urine of patients with inherited kidney disease has been reported, it remains low

Method used

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  • Establishment method of urine cell induced pluripotent stem cells of acute kidney injury patient
  • Establishment method of urine cell induced pluripotent stem cells of acute kidney injury patient
  • Establishment method of urine cell induced pluripotent stem cells of acute kidney injury patient

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Embodiment Construction

[0065] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0066] according to Figure 1 to Figure 8 As shown, a method for establishing induced pluripotent stem cells from urine cells of patients with acute kidney injury is as follows:

[0067] Isolation and culture of urine kidney cells from patients with acute kidney injury;

[0068] Such as figure 1 As shown in , use a special device such as a sterile centrifuge tube to collect patients in the recovery period of acute kidney injury. The mid-morning urine is abou...

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Abstract

The invention provides an establishment method of urine cell induced pluripotent stem cells of an acute kidney injury patient. The establishment method comprises the following steps of step 1, separating and culturing urine kidney cells A of the patient with acute kidney injury, analyzing the karyotype of the cells, and identifying the specific molecular expression of the kidney by immunofluorescence; step 2, transferring expression vectors TetO-FUW-OSKM of four transcription factors of OCT4, SOX2, KLF4 and c-MYC of a mouse source and an activation expression vector FUW-M2rtTA into the urine kidney cells A through a nuclear transfection technology to obtain urine kidney cells B; and step 3, culturing the urine kidney cells B, adding doxycycline hydrochloride into a culture solution of the urine kidney cells B for induction, establishing a urine cell induced pluripotent stem cell line, and performing immunofluorescence identification on pluripotent molecules of the cell line, therefore, according to the method, the urine kidney cells of the acute kidney injury patient can be separated, and induced pluripotent stem cells are established.

Description

technical field [0001] The invention belongs to the technical field of establishing induced pluripotent stem cells, in particular to a method for establishing induced pluripotent stem cells in urine cells of patients with acute kidney injury. Background technique [0002] In recent years, with the development of regenerative medicine, the research and application of stem cells and other related technologies in the medical field have attracted increasing attention, and the establishment of induced pluripotent stem cells (iPSCs) derived from adult cells has made Individualized medicine for patients has become possible, and it provides a new idea for using iPSCs to rescue kidney injury. In vitro isolation of urine cells is currently the best way to non-invasively obtain adult cells from patients. The kidneys of a healthy adult filter about 100 liters of body fluid every day, only 2 liters are excreted normally with urine, and about 2000-7000 exfoliated cells exist in the urine...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N5/10C12N15/85
CPCC12N5/0696C12N15/85C12N2501/119C12N2501/155C12N2501/11C12N2501/91C12N2501/16C12N2501/998C12N2500/32C12N2500/44C12N2501/33C12N2501/115C12N2501/235C12N2500/38C12N2501/602C12N2501/603C12N2501/604C12N2501/606C12N2500/30C12N2506/25C12N2800/107C12N2510/00
Inventor 金永张曼玲李荣凤曹长春
Owner NANJING MEDICAL UNIV
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