DNA measurement showed that approximately 99% of DNA was removed by 1 cycle of the decellularization process when compared with fresh intestinal tissue (2.9 ± 0.3 ng/mg vs. 194.2 ± 12.0, P < 0.0001), with no significant difference between 1 and 4 cycles (Fig. 2A).
The completion of the decellularization process was confirmed by histology (hematoxylin and eosin (H&E) staining) and DNA assay.
The decellularization process decreased the mechanical strength by 19.42% (n = 10, P < 0.05) in DECBM compared to NBM.
Hematoxylin and eosin staining showed a preserved ECM structure at the end of the decellularization process with no residual cells, confirmed by DAPI fluorescent staining.
The duration of the decellularization process for rat calvaria was optimized by 0.5% SDS and 0.1% NH4OH dissolved in distilled water [ 38, 39].
The decellularization process effectively removed the nucleic acids, as shown by the absence of fluorescence from all DC-ECMs stained with PI Supporting Information Figuree S2).
Methods to decellularize tissue and the criteria used to evaluate the efficacy of the decellularization process have been well established for porcine, rat and mouse brains9,10,12,14,30,48.
(b) SDS-PAGE elucidated the electrophoretic mobility and banding pattern of decellularized bECM favoring high molecular weight proteins, and the reproducibility of the decellularization process on the postnatal rat brain tissue across 4 litters (n = 3 brains/litter).
Baiguera et al.12 reported that elastin and sulfated glycosaminoglycans were retained following the decellularization process.
