Inves tigating The Presence of Extracellular Mitochondriain The Content of The S tem Cells from HumanExfoliated Deciduous Teeth (SHED) in Normal CultureConditions

Objective: Mitochondria are dynamic organelles that can enterthe extracellular space under certain conditions as signalingorganelles. Mitochondrial release from s tressed cells has beendebated for years, while the presence of mitochondria in theextracellular space of cells under normal culture conditions isunknown. S tem cells from human exfoliated deciduous teeth(SHED) and their secretome, due to their ease of access andwide therapeutic applications, have been regarded as a specificcandidate for cell therapies. Therefore, the present s tudy aimedto inves tigate the presence of extracellular vesicles containingmitochondria or naked mitochondria in a culture medium ofSHED in normal culture conditions.Materials and Methods: SHED with a density of ۴۵ ×۱۰۴ cell/cm۲ were cultured in DMEM supplemented with ۱۰% FBS. Afterthe cells reached ۹۰% confluency, conditioned media werecleared from cellular debris by centrifuging at ۴۰۰ g for ۱۰ minutes.Mitochondria isolation was performed throughout ۱۰۰۰۰g, ۱۰ minutes, at ۴ °C. The pellet was solved in PBS (free calciumand magnesium) and prepared for transmission electron microscopy(TEM) and Dynamic Light Scattering (DLS) analysis.Results: We demons trate that cells could release mitochondriaunder normal culture conditions. The presence, morphology,and size of mitochondrial-related particles were detected withTEM and DLS.Conclusion: Based on the TEM and DLS results, in normalculture conditions, mitochondria are released from the cells inthe form of naked, vesicular, and even swollen mitochondria

کلیدواژه ها:

Extracellular Vesicles ، Mesenchymal S tem Cells ، Mitochondria