Decellularized Kidney Scaffolds Research Articles

Effect of Co-culture of mesenchymal stem cell and glomerulus endothelial cell to promote endothelialization under optimized perfusion flow rate in whole renal ECM scaffold.

In recent era, many researches on implantable bio-artificial organs has been increased owing to large gap between donors and receivers. Comprehensive organ based researches on perfusion culture for cell injury using different flow rate have not been conducted at the cellular level. The present study investigated the co-culture of rat glomerulus endothelial cell (rGEC) and rat bone marrow mesenchymal stem cells (rBMSC) to develop micro vascularization in the kidney scaffolds culturing by bioreactor system. To obtain kidney scaffold, extracted rat kidneys were decellularized by 1% sodium dodecyl sulfate (SDS), 1% triton X-100, and distilled water. Expanded rGECs were injected through decellularized kidney scaffold artery and cultured using bioreactor system. Vascular endothelial cells adhered and proliferated on the renal ECM scaffold in the bioreactor system for 3, 7 and 14 days. Static, 1​ml/min and 2​ml/min flow rates (FR) were tested and among them, 1​ml/min flow rate was selected based on cell viability, glomerulus character, inflammation/endothelialization proteins expression level. However, the flow injury was still existed on primary cell cultured at vessel in kidney scaffold. Therefore, co-culture of rGEC​+​rBMSC found suitable to possibly solve this problem and resulted increased cell proliferation and micro-vascularization in the glomerulus, reducing inflammation and cell death which induced by flow injury. The optimized perfusion rate under rGEC​+​rBMSC co-culture conditions resulted in enhanced endocellularization to make ECM derived implantable renal scaffold and might be useful as a way of treatment of the acute renal failure.

Naphthalenephenylalanine-phenylalanine-glycine-arginine-glycine-aspartic promotes self-assembly of nephron progenitor cells in decellularized scaffolds to construct bioengineered kidneys

The shortage of donor kidneys is an important factor restricting kidney transplantation for patients with end-stage renal disease. To overcome this problem, we used decellularized kidney scaffolds and nephron progenitor cells (NPCs) as seed cells to construct bioengineered kidneys (BEKs). To reduce the effect of extracellular matrix (ECM) loss during the decellularization process on the cell growth microenvironment, we used dextrose to minimize collagen loss in decellularized kidney scaffolds. At the same time, to further improve the growth microenvironment of seed cells in the decellularized scaffolds, we modified the decellularized scaffolds with the self-assembling polypeptide Naphthalenephenylalanine-phenylalanine-glycine-arginine-glycine-aspartic (Nap-FFGRGD) to promote the adhesion and proliferation of seed cells in the scaffolds. NPCs were perfused into the decellularized kidney scaffolds and then the BEKs were cultured in vitro and transplanted in vivo. Markers of podocytes and renal tubules expressed in the glomeruli and renal tubules of the BEKs were detected by immunofluorescence staining, respectively were, suggesting that NPCs can continue to differentiate into renal cells and achieve nephron segment-specific re-population through self-assembly. These results indicate that by relying on the microenvironment provided by Nap-FFGRGD modified decellularized scaffolds, NPCs can be used to construct BEKs for transplantation in the future due to the self-assembly properties of organoids.

Kidney bioengineering by using decellularized kidney scaffold and renal progenitor cells

Patients with end-stage renal disease often need dialysis to maintain their lives because of donor organ shortage. The creation of a transplantable graft to permanently replace kidney function would overcome the organ shortage problem and the morbidity associated with immunosuppression. In the present study, we decellularized rat kidneys by the perfusion of detergent, yielding acellular scaffolds with the vascular, uretic, as well as cortical and medullary architecture. To regenerate the functional organ, we seeded tubular epithelial cells and mouse kidney progenitor cells from the ureter together with endothelial cells and mouse kidney progenitor cells from the renal artery. The renal constructs from seeded cells were cultured in a whole-organ bioreactor. After 3 months of organ culture, the seeded cells formed renal tubules, grew in the glomeruli, and some mouse kidney progenitor cells were also scattered in the interstitium. We tested the function of the bioengineered kidney with standardized perfusate in vitro. The bioengineered kidney not only produced urine but also reabsorbed albumin, glucose, and calcium. We conclude that seeded cell-based bioengineering of kidneys with physiological secreting and reabsorbing properties is possible and holds therapeutic promise.

Successful Male Rabbit Whole Kidney Decellularization For Tissue Engineering

Abstract Background Renal transplantation represents the only cure for end stage renal disease. Shortage of donated organs, immune compatibility and post-transplantation complications urges the search for another radical and revolutionary treatment . Whole kidney decellularization is a promising technique, hopefully offering a threedimensional natural kidney scaffold for engineering a patient’s compatible kidney after recellularization with patient’s own cells. The current study aimed at successfully decellularizing the male rabbit kidney while preserving the histological profile of the kidney decellularized extracellular matrix. Materials and Methods Kidneys of ten male New Zealand White Rabbits weighing (1000- 1500 gm) were harvested and sorted into two groups. Control Group I included the ten right kidneys. Decellularization Group II included the ten left kidneys; harvested carefully and kept frozen until decellularization. They were decellularized using 0.5% sodium dodecyl sulfate for 5-6 hours. Kidneys from both groups were processed similarly for histological examination. Results Decellularized kidney Scaffolds showed complete removal of cellular materials. Meanwhile, the structural profile of the decellularized kidney scaffolds was apparently well preserved. Conclusion The current study yielded an efficient decellularization of male rabbit kidney paving the way for future kidney recellularization.

Heparin modification improves the re-endothelialization and angiogenesis of decellularized kidney scaffolds through antithrombosis and anti-inflammation in vivo.

BackgroundConstructing tissue-engineered kidneys using decellularized kidney scaffolds (DKS) has attracted widespread attention as it is expected to be the key to solving the shortage of donor kidneys. However, thrombosis and the host inflammatory response are unfavorable factors that hider the re-endothelialization and vascularization of the decellularized scaffolds.MethodsHeparin was immobilized into the DKS using the method of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide/N-hydroxysuccinimide (EDC/NHS) activation. Fourier-transform infrared (FTIR) spectra were used to verify the heparinization of DKS. Human umbilical vein endothelial cells (HUVECs) were seeded and cultured in the DKS, then the sliced scaffolds were transplanted subcutaneously into nude mouse. Scanning electron microscopy and a series of histochemical stains including hematoxylin and eosin (H&E), elastic Verhöeff-Van Gieson (EVG), Sirius red, Masson’s trichrome, and toluidine blue (TB) staining were used for morphological characterization. The qRT-PCR analysis, immunohistochemistry (IHC), and immunofluorescence (IF) staining were used to determine the expression of related molecular markers.ResultsThe rat DKS completely retained the extracellular matrix and heparinized modification. The H&E staining results showed there were more HUVECs covering the internal surfaces of tubular structures in the HEP-DKS group compared with the DKS group. The IF analysis results revealed that CD31, Ki67, and CD206 had higher positive rates in HUVECs in the HEP-DKS group compared to the DKS group. Both groups of scaffolds showed blood vessel formation via H&E staining, and there were more blood vessels in the HEP-DKS group compared with the native DKS group (P<0.05). The qRT-PCR results showed that the levels of IL-1β, IL-6, and TNF-α in the HEP-DKS group were significantly lower than those of the native DKS group, while the expression level of IL-10 was significantly higher than that in the native DKS group (P<0.05).ConclusionsHeparin modification improves the re-endothelialization and vascular regeneration of the DKS through anticoagulation in vitro and in vivo. The anti-inflammatory effect of heparin on the transplanted host was initially confirmed, and it is considered that this effect may play a non-negligible role in promoting DKS re-endothelialization and angiogenesis. Heparinized DKS is therefore a promising candidate for kidney tissue engineering.

Decellularized kidney scaffold alters the healing response in chronic renal failure.

Decellularized (DC) kidney scaffold shows great potential for renal recovering. Our study explored the effect of the DC kidney scaffolds treating on chronic renal failure (CRF) through grafting them on 5/6 nephrectomized (5/6 Nx) rat kidneys compared with gelatin sponges covered the incision edges. Blood urea nitrogen and angiotensin II were significantly lower in most time in scaffold-grafted groups. Remnant kidney tumor necrosis factor-α and fibroblast growth factor in scaffold-grafted groups significantly reduced in majority of time points compared with controls. But platelet-derived growth factor-BB showed a different varied tendency, first higher in scaffold groups on week 2, 4, 6, but lower on week 8, finally no difference on week 12 compared with gelatin-sponge groups. In addition, the index of glomerular sclerosis was significantly lesser in scaffold-grafted groups, and, the accumulation of collagen III and collagen IV decreased in scaffold-grafted groups on week 6, 8, 12 compared with gelatin-sponge groups. Moreover, DC scaffolds enhanced the expression of CD133 on week 2, 6, 8, 12. In conclusion, DC kidney scaffold altered the healing response after 5/6 nephrectomy and ameliorated renal injury to some degree. Therefore, DC kidney scaffold could be a promising therapeutic method on CRF.

Effect of heparinization on promoting angiogenesis of decellularized kidney scaffolds.

Native decellularized extracellular matrix provides an adequate platform for tissues and organs and promotes the development of organogenesis and tissue remodeling. However, thrombosis poses a great challenge that hinders the transplantation for a substantial organ in vivo. Therefore, anticoagulation and re-reendothelialization of organ biological scaffolds are the primary concerns to be addressed before orthotopic transplantation. Herein, a heparinized decellularized kidney scaffold (HEP-DKSs) was prepared using end-point attachment technology, followed by binding the vascular endothelial growth factor (VEGF) to greatly improve the hemocompatibility and angiogenesis of DKSs. Based on the anticoagulant, co-culture of human umbilical vein endothelial cells, and subcapsular transplantation of kidney experiments, HEP-VEGF-DKSs are shown to reduce platelet adhesion, which is crucial for subsequent vascularization and slow release of heparin and VEGF, suggesting its ability of improve neovascularization. Taken together, these data indicated an optimal anticoagulation function of HEP-VEGF-DKSs and the potential of vascularization for regeneration of whole decellularized kidney.

Preparation of Decellularized Kidney Scaffolds in Rats.

Tissue engineering is a cutting-edge discipline in biomedicine. Cell culture techniques can be applied for regeneration of functional tissues and organs to replace diseased or damaged organs. Scaffolds are needed to facilitate the generation of three-dimensional organs or tissues using differentiated stem cells in vivo. In this report, we describe a novel method for developing vascularized scaffolds using decellularized rat kidneys. Eight-week-old Sprague-Dawley rats were used in this study, and heparin was injected into the heart to facilitate flow into the renal vessels, allowing heparin to perfuse into the renal vessels. The abdominal cavity was opened, and the left kidney was collected. The collected kidneys were perfused for 9 h using detergents, such as Triton X-100 and sodium dodecyl sulfate, to decellularize the tissue. Decellularized kidney scaffolds were then gently washed with 1% penicillin/streptomycin and heparin to remove cellular debris and chemical residues. Transplantation of stem cells with the decellularized vascular scaffolds is expected to facilitate the generation of new organs. Thus, the vascularized scaffolds may provide a foundation for tissue engineering of organ grafts in the future.

Preparation of Decellularized Kidney Scaffolds in Rats

Preparation of Decellularized Kidney Scaffolds in Rats

Visualization and Evaluation of Chemoembolization on 3D Decellularized Kidney Scaffold

Trans-arterial chemoembolization (TACE) has emerged as an effective palliative treatment for patients suffering from renal cell carcinoma. However, the current in vitro models for evaluating embolic performance and drug release kinetics of drug loaded embolic agents are limited since they greatly simplified the in vivo vascular systems as well as the three-dimensional (3D) structure of the organs. In this study, we developed a translucent decellularized rat kidney scaffold which could be applied on the evaluation of liquid based emulsions for TACE treatment. The characterization result of the decellularized kidney scaffold showed efficient decellularization as well as the integrity of the extracellular matrix with an enhanced transmittance up to 3.7% of the whole organ model. Using this model, we evaluated the clinically used doxorubicin-loaded ethiodised oil based emulsion with and without an emulsifier such as hydrogenated caster oil-40 (HCO-40). We found that the emulsion with enhanced stability could achieve comparably sustained drug release and perform better occlusion level especially when a mimicking accessory blood supply was available. Such a model that possesses intricate vascular networks and abundant in vivo physiological information might provide a unique approach for spatiotemporally evaluating the embolic performance and drug release kinetics of chemotherapeutic embolic agents and thus advance the development of novel embolic agents.

Kidney tissue engineering using a well-preserved acellular rat kidney scaffold and mesenchymal stem cells.

The aim of this study was to acquire an effective method for preparation of rat decellularized kidney scaffolds capable of supporting proliferation and differentiation of human adipose tissue derived mesenchymal stem cells (AD-MSCs) into kidney cells. We compared two detergents, the sodium dodecyl sulfate (SDS) and triton X-100 for decellularization. The efficiency of these methods was assessed by Hematoxylin and Eosin (H&E), 4', 6 diamidino-2-phenylindole and immunohistochemistry (IHC) staining. In the next step, AD-MSCs were seeded into the SDS-treated scaffolds and assessed after three weeks of culture. Proliferation and differentiation of AD-MSCs into kidney-specific cell types were then analyzed by H&E and IHC staining. The histological examinations revealed that SDS was more efficient in removing kidney cells at all-time points compared to triton X-100. Also, in the SDS-treated sections the native extracellular matrix was more preserved than the triton-treated samples. Laminin was completely preserved during decellularization procedure using SDS. Cell attachment in the renal scaffold was observed after recellularization. Furthermore, differentiation of AD-MSCs into epithelial and endothelial cells was confirmed by expression of Na-K ATPase and vascular endothelial growth factor receptor 2 (VEGFR-2) in seeded rat renal scaffolds, respectively. Our findings illustrated that SDS was more effective for decellularization of rat kidney compared to triton X-100. We presented an optimized method for decellularization and recellularization of rat kidneys to create functional renal natural scaffolds. These natural scaffolds supported the growth of AD-MSCs and could also induce differentiation of these cells into epithelial and endothelial cells.

Insights Image for "Human ucMSCs seeded in a decellularized kidney scaffold attenuate renal fibrosis by reducing epithelial-mesenchymal transition via the TGF-β/Smad signaling pathway".

Insights Image for "Human ucMSCs seeded in a decellularized kidney scaffold attenuate renal fibrosis by reducing epithelial-mesenchymal transition via the TGF-β/Smad signaling pathway".

Effect of Heparinization on Promoting Angiogenesis of Decellularized Kidney Scaffolds

In recent years, the native decellularized extracellular matrix (ECM) provides a scaffold for tissues and organs, but also supports organogenesis and tissue remodeling. However, the anti-coagulation properties have always been a challenge in the creation of the ideal three dimensional bioscaffold materials for tissue engineering organ construction. This hurdle is also an inevitable issue that researchers have to face in the revascularization of the decellularized kidney scaffolds (DKSs). This manuscript describes the strategies to accelerate the endothelialization of blood vessels of DKSs based on anticoagulation, compared to previous reports in our research. In this study, we prepared a heparinized decellularized kidney scaffold (Hep-DKSs) by end-point attachment (EPA) technology, then binding the vascular endothelial growth factor (VEGF) with the Hep-DKSs in order to rapidly promote the formation of endothelialization of DKSs. Following, the anticoagulant properties and cytocompatibility of the modified scaffolds were investigated in vitro by means of Toluidine blue staining, Platelet adhesion assay and Co-culture of human umbilical vein endothelial cells (HUVECs). Subsequently, the modified scaffolds were transplantated to intrarenal subcapsular of the recipient rat. Then the transplanted specimens were observed on 1, 3, 7, 14, 21 and 28 days after transplantation. The results in vivo showed that the numbers of neovascularization were higher in the heparin cross-linking VEGF decellularized kidney scaffolds (HEP-VEGF-DKSs) than that of other decellularized renal scaffolds. Taken together, these data suggest that HEP-VEGF-DKSs have a good anticoagulation property and biocompatibility, and can slowly release heparin and VEGF, suggesting its ability of improving neovascularization better and faster and potential use for regeneration of whole decellularized kidney.

Human ucMSCs seeded in a decellularized kidney scaffold attenuate renal fibrosis by reducing epithelial-mesenchymal transition via the TGF-β/Smad signaling pathway.

Renal fibrosis occurs largely through epithelial-mesenchymal transition (EMT). This study explored the beneficial effects of a human umbilical cord mesenchymal stem cell-loaded decellularized kidney scaffold (ucMSC-DKS) on renal fibrosis in a rodent model of post-transplantation renal failure, and the underlying mechanism. Rat-derived DKSs were examined after preparation, and then recellularized with human ucMSCs to prepare cell-loaded patches. A rat model of renal failure was established after subtotal nephrectomy (STN). The cell patches were transplanted to remnant kidneys. Changes in renal function, histology, EMT, and proteins related to the transforming growth factor-β (TGF-β)/Smad signaling pathway in the remnant kidneys were examined 8 weeks after surgery, compared with non-cell patch controls. The DKSs were acellular and porous, with rich cytokine and major extracellular matrix components. The ucMSCs were distributed uniformly in the DKSs. Renal function was improved, renal fibrosis and EMT were reduced, and the TGF-β/Smad signaling pathway was inhibited compared with controls at 8 weeks after ucMSC-DKS patch transplantation. The ucMSC-DKS restores renal function and reduces fibrosis by reducing EMT via the TGF-β/Smad signaling pathway in rats that have undergone STN. It provides an alternative for renal fibrosis treatment.

Induced Intermediate Mesoderm Combined with Decellularized Kidney Scaffolds for Functional Engineering Kidney.

Chronic kidney disease is a severe threat to human health with no ideal treatment strategy. Mature mammalian kidneys have a fixed number of nephrons, and regeneration is difficult once they are damaged. For this reason, developing an efficient approach to achieve kidney regeneration is necessary. The technology of the combination of decellularized kidney scaffolds with stem cells has emerged as a new strategy; however, in previous studies, the differentiation of stem cells in decellularized scaffolds was insufficient for functional kidney regeneration, and many problems remain. We used 0.5% sodium dodecyl sulfate (SDS) to produce rat kidney decellularized scaffolds, and induce adipose-derived stem cells (ADSCs) into intermediate mesoderm by adding Wnt agonist CHIR99021 and FGF9 in vitro. The characteristics of decellularized scaffolds and intermediate mesoderm induced from adipose-derived stem cells were identified. The scaffolds were recellularized with ADSCs and intermediate mesoderm cells through the renal artery and ureter. After cocultured for 10days, cells adhesion and differentiation was evaluated. Intermediate mesoderm cells were successfully induced from ADSCs and identified by immunofluorescence and Western blotting assays (OSR1 + , PAX2 +). Immunofluorescence showed that intermediate mesoderm cells differentiated into tubular-like (E-CAD + , GATA3 +) and podocyte-like (WT1 +) cells with higher differentiation efficiency than ADSCs in the decellularized scaffolds. Comparatively, this phenomenon was not observed in induced intermediate mesoderm cells cultured in vitro. In this study, we demonstrated that intermediate mesoderm cells could be induced from ADSCs and that they could differentiate well after cocultured with decellularized scaffolds.

Optimization of preparation of rat kidney decellularized scaffold by combining freeze-thawing with perfusion

We explored the improved method to prepare decellularized kidney scaffold and provide experimental basis for kidney tissue engineering and renal pathology and toxicology in vitro research. We perfused rat kidneys with PBS (group control) and prepared the decellularized kidney scaffolds with sodium dodecyl sulfate (SDS) (group S), Triton X-100 combined with SDS (group TS), and Triton X-100 combined with SDS after repeated freezing and thawing (group FTS) in different flow velocity. Meanwhile we measured their fluid distributions and vascular resistance. We examined the degree of decellularization of acellular scaffolds by HE, DAPI staining and DNA quantification. We examined the retention of main composition and structural integrity of decellularized scaffolds by Masson, PAS and immunohistochemical staining. We also detected the ultrastructure, cytotoxicity and the level of growth factor of the scaffolds by scanning electron microscope, MTT and ELISA, respectively. The results showed that the time of decellularization in group FTS was less than that in group S and TS. The vascular resistance of scaffolds decellularized at 10 mL/min flow velocity was lower. The fluid distribution in groups S, TS and FTS was different from that in control group. No residual cell was detected by HE and DAPI staining. DNA content was less than 50 ng/mg. Masson, PAS and immunohistochemical staining results showed that there was extracellular collagen, polysaccharide, type I collagen, type IV collagen, fibronectin and laminin in the decellularized scaffolds, and the scanning electron microscope result showed the scaffolds had the honeycomb structure. The cytotoxicity level of decellularized scaffolds was between grade 0 to 1. The level of VEGF, EGF, IGF-1 and PDGF-BB in group FTS were significantly higher than those in group S and TS. In concluding, combining freeze-thawing with perfusion can produce more ideal and effective whole organ decellularized scaffold of rat kidney, and make a foundation for the study of kidney tissue engineering and in vitro pathology and toxicology of kidney.

Immobilization of heparin on decellularized kidney scaffold to construct microenvironment for antithrombosis and inducing reendothelialization.

In recent years, rapid development of tissue engineering technology provides possibilities for the construction of artificial tissues or organs. In construction of engineered kidneys, researchers used native decellularized extracellular matrix (ECM) as the scaffolds to recellularization. However, thrombosis has been a great issue that hinders the progress of transplantation in vivo. In this study, heparin was immobilized to the collagen part of decellularized scaffold with collagen-binding peptide (CBP). Through the anticoagulant and endothelial cell reperfusion experiments, it can be demonstrated that the heparinized scaffolds absorbed less platelets and red blood cells which can effectively reduce the formation of thrombosis. Moreover, it is conducive to long-term adhesion of endothelial cells which is important for the formation of subsequent vascularization. Taken together, our results reveal that the whole kidney can be modified by CBP-heparin composite to reduce the thrombosis and provide the better conditions for neovascularization.

Biocompatibility and hemocompatibility of efficiently decellularized whole porcine kidney for tissue engineering.

Whole kidney decellularization is a promising approach in regenerative medicine for engineering a functional organ. The reaction of the potential host depends on the biocompatibility of these decellularized constructs. Despite the proven ability of decellularized kidney scaffolds to guide cell attachment and growth, little is known about biocompatibility and hemocompatibility of these scaffolds. Our aim is to prepare decellularized kidneys of a clinically relevant size and evaluate its biocompatibility and hemocompatibility. Porcine kidneys were cannulated via the renal artery, and then perfused with 0.1% sodium dodecyl sulfate solution. Hematoxylin and eosin as well as DAPI staining confirmed cellular clearance from native kidneys in addition to preservation of the microstructure. SEM confirmed the absence of any cellular content within the scaffold, which is maintained in a well-organized 3D architecture. Decellularized kidneys retained the intact renal vasculature upon examination with contrast radiography. The essential structural extracellular matrix molecules were well-preserved. Scaffolds were susceptible to enzymatic degradation upon collagenase treatment. Scaffolds showed a good hemocompatibility when exposed to porcine blood. Decellularization was efficient to remove 97.7% of DNA from native kidneys in addition to the immunogenic and pathogenic antigens. Scaffolds did not induce the human immune response in vitro. Decellularized kidneys were non-cytotoxic to pig kidney cells (PKs). PKs were able to grow and proliferate within the decellularized renal scaffolds with maintaining a higher function than cells grown as monolayers. Thus, we have developed a rapid decellularization technique for generating biocompatible kidney scaffolds that represents a step toward development of a transplantable organ. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2034-2047, 2018.

Development of an experimental model of a decellularized kidney scaffold by perfusion mode and analyzing the three-dimensional extracellular matrix architecture by edge detection method

Renal transplant is treatment of choice for the patients with end stage renal disease. The kidney transplants are expensive and there are risks of immunological and infectious complications. We planned to develop an in vitro decellularized kidney scaffold model using sheep kidney. Kidney decellularization was carried out by perfusing chemical detergents such as sodium dodecyl sulfate (SDS), SDS and trypsin, and SDS and ethylenediaminetetraacetic acid solvent solution. Complete kidney was decellularized in 5 days by perfusing various chemical detergents in time-dependent intervals. Histological finding revealed the complete removal of cellular material in various regions of renal corpuscle, distal convoluted tubules, other cortex and medulla region. Details of interlobular veins and arteries were seen through naked eyes after trypan blue dye injection. We used edge detection technique for developing a three-dimensional (3-D) image (Image J software) for nephrological vasculature constructed of decellularized kidney scaffold specimen. This technique opens a gateway for the whole organ decellularization by perfusion technology and further imaging of its 3-D extracellular matrix texture by edge detection technique software.

Streamlined System for Conducting In Vitro Studies Using Decellularized Kidney Scaffolds.

Kidney regeneration is a complex process that can only be studied in vitro at a limited capacity due to the inherent structural and functional complexity of its tissues. Thus, a suitable platform for conducting cellular response and development should be established from decellularized tissues with intact microarchitecture. In this study, a modular streamlined system was developed to allow manageable handling and setup of in vitro studies using decellularized rat kidneys. The system is composed of commercially available parts that can be reused, interchanged, and reconfigured based on the desired experimental stage and process. Decellularization was confirmed through time-lapse observation, stained tissue sections, genetic material quantification, and protein analysis. The capacity of the bioreactor design to support cell-seeded decellularized kidney constructs was tested by determining viability of seeded podocytes and endothelial cells. Based on the results, decellularized kidneys with renal proteins and intact microstructures can be achieved in relatively shorter periods compared (12 h) to established protocols (96-120 h). The minimalistic kidney bioreactor design not only maintained sterility of decellularized kidney without cells but also permitted manageable maintenance of cell-immobilized constructs for up to 1 week. Through this streamlined system, sustainable and reproducible in vitro experiments for kidney regeneration can be designed and conducted using decellularized kidney as a platform for cell growth and development.