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Objective: Bone marrow cell extract (BMCE) was previously reported to restore salivary gland hypofunction caused by irradiation injury. Proteins were shown to be the main active factors in BMCE. However, BMCE therapy requires multiple injections and protein denaturation is a concern during BMCE storage. This study aimed to preserve, by lyophilization (freeze‐drying), the bioactive factors in BMCE. Methods: We developed a method to freeze‐dry BMCE and then to analyze its ingredients and functions in vivo. Freeze‐dried (FD) BMCE, freshly prepared BMCE (positive control), or saline (vehicle control) was injected into the tail vein of mice that had received irradiation to damage their salivary glands. Results: Results demonstrated that the presence of angiogenesis‐related factors and cytokines in FD‐BMCE remained comparable to those found in fresh BMCE. Both fresh and FD‐BMCE restored comparably saliva secretion, increased cell proliferation, upregulated regenerative/repair genes, protected salivary acinar cells, parasympathetic nerves, and blood vessels from irradiation‐damaged salivary glands. Conclusion: Lyophilization of BMCE maintained its bioactivity and therapeutic effect on irradiation‐injured salivary glands. The advantages of freeze‐drying BMCE are its storage and transport at ambient temperature. [ABSTRACT FROM AUTHOR]

Objective A challenge in studying human salivary glands is to maintain the cells ex vivo in their three-dimensional (3D) morphology with an intact native extracellular matrix ( ECM) environment. This paper established a human salivary 3D organotypic slice culture model that could maintain its physiological functions as well as allowing a direct visualization of the cells. Methods Human salivary biopsies from six patients were embedded in agarose and submerged in cold buffer for thin (50 μm) sectioning using a vibratome. 'Salivary slices' were mechanically supported by a porous membrane insert that allowed an air-liquid interface and cultured in serum-free culture media. Cell viability, proliferation, apoptosis, physiological functions, and gene expression were assessed during 14 days of culture. Results Human salivary slices maintained cell survival (70-40%) and proliferation (6-17%) for 14 days ex vivo. The protein secretory (amylase) function decreased, but fluid (intracellular calcium mobilization) function was maintained. Acinar, ductal, and myoepithelial cell populations survived and maintained their 3D organization within the slice culture model. Conclusion The human salivary slice culture model kept cells alive ex vivo for 14 days as well as maintaining their 3D morphology and physiological functions. [ABSTRACT FROM AUTHOR]