Abstract:

Objective　To produce the decellularized biological renal scaffold of the pig, New Zealand white rabbit and SD rat, and then to explore the influence of porcine, leporine and murine renal biological scaffolds on the co-cultured seed cells HEK.　Methods　Adult healthy kidneys from 10 pigs, 28 New Zealand white rabbits, and 28 SD rats were harvested and divided into normal groups and scaffold groups randomly and equally. The scaffold group kidneys were perfused for decellularization with heparinized PBS, 1% Triton X-100 and 1% SDS through the renal artery successively. The kidneys in each group were characterized in histomorphology and detected for mechanical properties. And the co-culture experiments were conducted using the slices of each group and HEK cells, and the expression of PCNA in HEK cells was analyzed by immunofluorescence assay using the value of the gray level.　Results　In each scaffold group after decellularization, the HE staining of cell nucleus in the slices was negative, Masson's trichrome staining of collagen was positive, and immunofluorescence staining of Collagen I and Collagen IV was also positive. And with the scanning electron microscopy, the faveolate orifice structure and the typical glomerulus-like structure were observed. Characterized by the mechanical properties, the Young's modulus of each scaffold group is similar with that of the corresponding normal group. After the co-culture, the PCNA/DAPI value of every renal scaffold group is higher than that of the blank control. Furthermore, there was no significant difference on the PCNA/DAPI of HEK between the pig, New Zealand white rabbit and SD rat renal scaffolds.  Conclusions　The decellularization method of our study can eliminate the cells in the porcine, leporine and murine kidney, retain the extracellular matrices and preserve the spatial structure and similar mechanical strength. And it is an effective way to produce the decellularized pig, New Zealand white rabbit and SD rat renal biological scaffolds. Importantly, the produced decellularized porcine, leporine and murine kidney scaffold slices can improve the proliferation activity of heterogeneous HEK cells when co-cultured in vitro, and there is no species difference on the improvement between the pig, New Zealand white rabbit and SD rat.

Key words:  , Decellularization, 　Biological scaffold, 　Kidney regeneration, 　Co-cuture, 　Tissue engineering