To characterize the quality of decellularization, the tissues before and after decellularization were fixed in 4% PFA for 30 min, immersed in 15% and then 30% sucrose solution, and embedded in OCT for cryosection. H&E staining was performed according to the manufacture protocol (Solarbio, Cat#G1121).
SDS‐PAGE was performed to qualitatively examine ECM protein content. Briefly, a small piece of native porcine kidney tissue was lysed in RIPA buffer. Digested ECM hydrogel was used because the decellularized ECM did not dissolve in common lysis buffers. The protein concentration of each sample was determined by BCA assay. Proteins were separated by 10% SDS‐PAGE, stained by Coomassie Blue, digested by trypsin and identified by LC‐MS/MS as previously described [31 (link)].
DNA quantification was performed by PicoGreen assay following previous reports [10 (link)]. Freeze‐dried porcine kidney tissues before and after decellularization were digested in Proteinase K overnight. DNA was quantified by the PicoGreen dsDNA Kit (Yeasen, Cat#12641ES01). Sulfated glycosaminoglycan (GAG) was quantified by the DMMB assay following the previous report [10 (link)].
SDS‐PAGE was performed to qualitatively examine ECM protein content. Briefly, a small piece of native porcine kidney tissue was lysed in RIPA buffer. Digested ECM hydrogel was used because the decellularized ECM did not dissolve in common lysis buffers. The protein concentration of each sample was determined by BCA assay. Proteins were separated by 10% SDS‐PAGE, stained by Coomassie Blue, digested by trypsin and identified by LC‐MS/MS as previously described [31 (link)].
DNA quantification was performed by PicoGreen assay following previous reports [10 (link)]. Freeze‐dried porcine kidney tissues before and after decellularization were digested in Proteinase K overnight. DNA was quantified by the PicoGreen dsDNA Kit (Yeasen, Cat#12641ES01). Sulfated glycosaminoglycan (GAG) was quantified by the DMMB assay following the previous report [10 (link)].
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