Application of human amniotic epithelial stem cells in preparation of medicine for treating cisplatin-induced acute kidney injury
A technology of acute kidney injury and stem cell preparations, applied in the field of biomedicine, can solve the problems of unassessed effects, insufficient number of patients recruited, etc., reduce the requirements for transplant matching, reduce the source of immune cells, and enhance the proliferation ability Effect
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Embodiment 1
[0061] This example provides a method for the isolation and culture of primary human amniotic epithelial stem cells. The specific materials and processes are as follows:
[0062] 1. Source of human amniotic membrane
[0063] In order to avoid microbial contamination of the birth canal, fetal placenta delivered by cesarean section was selected. Due to the stimulation of labor signals after term, the amniotic membrane will undergo apoptosis, so it is advisable to use premature fetal placenta (before 38 weeks). After the puerpera signed the informed consent, the placental tissue of the healthy puerpera (HIV, syphilis, hepatitis A, hepatitis B, hepatitis C and other serological reactions were all negative) after cesarean section was taken, the placenta was cut with a cross knife, and the whole amniotic membrane was obtained by mechanical separation.
[0064] 2. Isolation of hAESCs (aseptic operation is required throughout the process)
[0065] (1) Obtain the placenta of infants ...
Embodiment 2
[0078] In this example, a cisplatin-AKI mouse model was constructed and processed in groups. The specific process is as follows:
[0079] 1. Preparation of cisplatin solution: Dissolve cisplatin powder in physiological saline, prepare a cisplatin solution with a concentration of 1mg / ml, and store it at -20°C.
[0080] 2. Mice preparation and grouping: 8-week-old C57BL / 6j male mice were purchased and fed adaptively in a barrier environment for one week, then randomly grouped according to body weight: normal control group, cisplatin treatment group (20mg / kg or 15mg / kg Cisplatin intraperitoneal injection) and hAESCs treatment group (hAESCs treatment the next day after cisplatin treatment). 9-10 mice per group.
[0081] 3. Cisplatin injection: the cisplatin-AKI model was prepared by intraperitoneal injection of cisplatin solution. The injection dose of cisplatin 20mg / kg is for severe AKI model, and 15mg / kg is for moderate AKI model.
[0082] 4. Stem cell injection: On the first...
Embodiment 3
[0085] This example verifies that hAESCs improve the survival rate of cisplatin-AKI mice, and the specific results are as follows:
[0086] Such as figure 1 As shown, the 6-day mortality rate of untreated mice was 100%. Using the severe cisplatin-AKI mouse model (20mg / kg cisplatin intraperitoneal injection), administration of hAESCs on the first day after cisplatin injection could make cisplatin - The mortality rate of AKI mice was reduced to about 50% (P<0.05).
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