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. 2022 Oct 7;10:1019437. doi: 10.3389/fbioe.2022.1019437

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Copyright © 2022 Xia, Yang, Hou, Wang, Li, Zhu and Fu.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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(A) Schematic representation of in vitro reprogramming of human nucleus pulposus (NP) cells by bulk electroporation of Forkhead-FOXF1). (B) Brightfield images of NP cells co-cultured in monolayers with PKH26-stained pCMV6 and FOXF1 extracellular vesicles (EVs, red), respectively, at 0, 2, and 7 d. (C) Representative images (bottom) of 4 × (top) and 10 × images (bottom) of cell-embedded three-dimensional (3D) agarose gels at days 0, 2, and 4 in culture (scale bar: 500 μm). Live cells are stained green (calcein-AM) and dead cells are stained red (ethidium-homodimer). (D) Quantified visual uptake by the percentage of cells with EVs over the total number of cells. (E) FOXF1 expression of FOXF1-EV-treated human NP cells in monolayers normalized to pCMV6-EV-treated cells. (F) Normalized to DNA content of autopsy and surgical human NP cells in 3D culture at 0, 2, and 4 weeks. Reproduced with permission from (Tang et al., 2021).