Fig. 1: Salivary tissue-organoid biobanking preserves primitive salivary cells.

A Schematic workflow outlining the steps involved in the processing of PG and SMG tissues to generate tissue-organoids, as well as their cryopreservation and biobanking from living or deceased male and female donors (created using Biorender.com). B Plot showing the age distribution among PG and SMG donors, considering both living and deceased individuals of both genders in the biobank. C) Plot depicting the weight distribution of PG and SMG tissues received at the laboratory, considering both living and deceased donors of both genders. D Plot illustrating the quantity of patient-derived tissue-organoid cryovials obtained from PG and SMG tissue masses received at the laboratory, considering both living and deceased donors of both genders. E Bar plot illustrating the distribution of PG and SMG tissue-organoids in the biobank, considering donors from both living and deceased individuals of both genders. F H&E staining of SMG and PG tissues and their corresponding tissue-organoids show characteristic ductal and acinar structures (100x magnification). G Representative brightfield images of cultured SMG and PG cells isolated from respective cryopreserved tissue-organoids (100x magnification). H Representative brightfield images of SMG and PG organoids cultured in 3D-matrigel (40x and 100x magnification) derived in culture following dissociation of respective tissue-organoids. I Representative immunofluorescence staining of 3D-matrigel-derived PG organoids with DAPI, Aquaporin 5, NKCC1, SMA, KRT5, KRT7, KRT8 and KRT14 antibodies. J Agarose gel (2%) image showing expression of AMY1A (162 bp), AQP5 (178 bp) in two consecutive 3D organoid cultures but not in 2D SMG and PG cultures by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). Gene expression was normalized to ACTB (494 bp). Uncropped gel images are supplied as Supplementary Fig. S12. K) Plot showing 3D OIC frequency from cryopreserved salivary gland tissue-organoids.