| Background: The incidence of chronic kidney disease(CKD) is increasing and the number of patients undergoing the end stage of chronic kidney disease is rising. At present, dialysis and kidney transplantation are still the main way to treat end-stage renal disease. Renal transplantation is the most effective treatment at the final stage of renal disease, but kidney shortage is a serious problem. So stem cell transplantation might be an ideal treatment method. In recent years, stem cells have attracted more and more attentions for their ability of differentiation. Stem cells have been used in tissue repair and regeneration and have become the focus of medical research. Embryonic stem cells(ESCs) can differentiate into various cell types including endoderm,ectoderm and mesoderm. Embryonic stem cells, therefore, can be seen as an effective tool for the treatment of kidney regeneration. We use gelatin microcryogels(GMs) carrying embryonic stem cells to research the protective effects of embryonic stem cells on rats with 5/6 nephrectomy and explore its mechanism in order to provide a certain experimental basis for the treatment of ESCs in kidney diseases.Objective: Construct the gelatin microcryogels-embryonic stem cells complexes; Detect the biological compatibility of the complexes; Build 5/6 nephrectomy rat model of chronic renal failure; Use the pedicled greater omentum flap packing embryonic stem cell-loaded gelatin microcryogels on 5/6 nephrectomized kidney to detect the renal protective effects on the rat model of CKD.Methods: (1) After gelatin microcryogels-embryonic stem cells complexes co-culture of 96h, the HE staining was performed to observe the cell adhesion and survival status; the cell proliferation was detected by CCK8; and the light microscope and two-photon fluorescence confocal imaging were also used to detect the viability of ESCs within GMs. (2) 40 male SD rats were randomly divided into Sham, only removal of renal capsule; NPX, 5/6 nephrectomy; NPX+OM, 5/6 nephrectomy and pedicled greater omental directly packing on remnant renal tissue; NPX+OM+ESCs, 5/6 nephrectomy and pedicled greater omental packing free ESCs on remnant renal tissue;NPX+OM+GMs+ESCs, 5/6 nephrectomy and pedicled greater omentum packing ESCs-loaded GMs on remnant renal tissue. Each experimental group comprised 8 rats.Rats were euthanized at weeks 12 after inducing CKD. Collecting blood samples and renal tissue samples, serum creatinine (SCr) and 24h urine protein was measured in all groups, PAS and MASSON staining were used to evaluate the renal pathology and renal tubular score, and a -SMA, TGF- ? COL-? immunohistochemistry and western blot analysis were used to evaluate the renal injury and repair situation.Results: (1) HE staining showed that ESCs-loaded GMs have uniform cell distribution,good cell attachment and viability; The light microscope and two-photon fluorescence confocal imaging showed that ESCs could be automatically loaded into GMs; CCK8 test showed that the GMs had no effect on the proliferation and growth of ESCs. (2)After 12 weeks, the pedicled greater omentum flap packing ESCs-loaded GMs on 5/6 nephrectomized rats had lower plasma creatinine levels; less glomerulosclerosis,tubulointerstitial injury compared with only 5/6 nephrectomized rats.Conclusions: Embryonic stem cells-loaded gelatin microcryogels have good biological compatibility. In the establishment rats model of 5/6 nephrectomy, it is proved that the pedicled greater omentum flap packing ESCs-loaded GMs on 5/6 nephrectomized rats group compared with only 5/6 nephrectomy rats group can effectively reduce the progression of chronic kidney disease and renal injury, antagonistic to renal fibrosis and show a sustained therapeutic effect. |
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