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. 2021 Jul;11(7):e384.
doi: 10.1002/ctm2.384.

Exosomes derived from human umbilical cord Wharton's jelly mesenchymal stem cells ameliorate experimental lymphedema

Zhao Ting  1   2 , Yan Zhi-Xin  3 , Tan You-Wen  4 , Yang Fu-Ji  1 , Sun Hui  1 , Mao Fei  1 , Zhu Wei  1 , Xu Wen-Rong  1 , Qian Hui  5 , Yan Yong-Min  5
Affiliations

Exosomes derived from human umbilical cord Wharton's jelly mesenchymal stem cells ameliorate experimental lymphedema

Zhao Ting et al. Clin Transl Med. 2021 Jul.
No abstract available

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Figures

FIGURE 1
FIGURE 1
MSC‐ex reduced lymphedema and enhanced lymphangiogenesis in vivo. (A) MSC markers of CD29, CD44, and CD206 detection by imaging flow cytometry. (B, C) Exosomal markers (CD9, CD63, CD81, TSG101) and non‐exosomal markers of Calnexin detection by imaging flow cytometry and western blot. (D) Representative images of MSC (20×). (E, F) Representative images of nano‐size vesicles photographed by transmission electron microscope (TEM) and atomic force microscope (AFM). Scale bar, 100 nm. (G) Nanoparticle tracking analyses of MSC‐ex in PBS. (H) Representative images of microsurgical ablation of major lymphatic conduits in the mouse (Hi). Schematic illustration of in vivo animal test for evaluating efficacy of MSC‐ex on lymphedema (Hii). (I) Distribution of Dir labeled MSC‐ex in mouse tail by in vivo fluorescent imaging 24 h post‐treatment. (J) Representative images of intradermal injection of methylene blue on day 42 resulted in transportation of the dye across incision site (n = 6). (K) Assessment of circumference directly at 0 cm, 1 cm, and 2 cm distal of incision site (n = 6; *P < 0.05 vs PBS, **P < 0.01 vs PBS). (L) Representative histology of mouse tail harvested on day 42. Dermal thickness was quantified (n = 6; **P < 0.01 vs PBS). Scale bar, 100 μm. (M) Representative images of immunohistochemical staining for LYVE‐1 positive lymphatics from mice treated with PBS, MSC or MSC‐ex on day 42. Number of LYVE‐1 positive lymphatics were quantified (n = 6; **P < .01 vs PBS). Scale bar, 100 μm. All data are presented as means ± SEM, Mann‐Whitney test. PBS, phosphate saline solution; MSCex, mesenchymal stem cells derived exosomes. LYVE‐1, lymphatic endothelial hyluronan receptor‐1
FIGURE 2
FIGURE 2
MSC‐ex exhibits dose‐dependent effect on lymphangiogenesis. (A) Representative fluorescent images of PKH26‐labeled MSC‐ex in HDLECs at 12, 24, and 36 h post MSC‐ex incubation. Scale bars, 10 μm. (B) CCK‐8 assay showing the effect of MSC‐ex on HDLEC proliferation at 24 h (n = 5; **P < .01 vs PBS). (C) Transwell migration assay of MSC‐ex treated HDLEC. (n = 5; **P < .01 vs PBS). Scale bars, 100 μm. (D) Matrigel tube formation of MSC‐ex treated HDLEC. (n = 4; *P < .05 vs PBS). Scale bars, 100 μm. (E) Representative immunofluorescent images of LYVE‐1 expression in PBS or MSC‐ex treated HDLECs at 24 h. Scale bars, 10 μm. (F) Western blot for LYVE‐1 in HDLEC treated with MSC‐ex at indicated time (n = 4; *P < .05 vs PBS). (G) Western blot for Ang2, Prox1, p‐Akt, and t‐Akt in MSC‐ex treated HDLEC at indicated time (n = 4; *P < .05 vs PBS). (H) Western blot for Ang1, Ang2, Prox1, and VEGFR3 in MSC‐ex or MSC from different batches. (I, J) Representative immunofluorescent images of Ang2 expression in HDLEC (n = 4; *P < .05 vs PBS, Scale bars, 10 μm.) and mouse tails (n = 4; *P < .05 vs PBS, Scale bars, 50 μm). All data are presented as means ± SEM, Mann‐Whitney test. HDLEC, human dermal lymphatic endothelial cells; MSC‐ex, mesenchymal stem cells derived exosomes; PBS, phosphate saline solution
FIGURE 3
FIGURE 3
Prolymphangiogenesis effect of MSC‐ex on HDLEC was mediated by Ang2. (A) Western blot for Ang2 in MSC‐ex from MSC transfected with Lenti‐Ang2 (MSC‐exAng2) (Ai), Ang2‐shRNA (MSC‐exshAng2) (Aii) (n = 4; *P < .05 vs MSC‐exGFP). (B) Transwell migration assay (n = 5; *P < .01 vs MSC‐exGFP, Scale bars, 200 μm) and matrigel tube formation of MSC‐exGFP/MSC‐exAng2 treated HDLEC (n = 4; *P < .05 vs MSC‐exGFP, Scale bars, 100 μm). (C) CCK‐8 assay of MSC‐exGFP/MSC‐exAng2 treated HDLEC (n = 4; *P < .05 vs MSC‐exGFP). (D) Transwell migration assay (n = 4; *P < .05 vs MSC‐exshCtr, Scale bars, 200 μm) and matrigel tube formation of MSC‐exshCtr/MSC‐exshAng2 treated HDLEC (n = 4; *P < .05 vs MSC‐exshCtr, Scale bars, 100 μm). (E) CCK‐8 assay of MSC‐exshCtr/MSC‐exshAng2 treated HDLEC (n = 4; *P < .05 vs MSC‐exshCtr). (F) Representative immunofluorescent images of Ang2 expression in MSC‐exshCtr/MSC‐exshAng2 treated mouse tails. Relative mean fluorescence intensity (rMFI) was quatntied (n = 4; *P < .05 vs MSC‐exshCtr). Scale bars, 10 μm. (G) Representative histology of mouse tail harvested on day 28. Dermal thickness was quantified on day 28 in tail skin (n = 4; *P < .05 vs MSCexshCtr). Scale bar, 100 μm. (H) Representative images of intradermal injection of methylene blue on day 28 (n = 4). (I) Assessment of the circumference directly on day 28 after PBS, MSC‐exshCtr, and MSCexshAng2 treatment (n = 4; *P < 0.05 vs MSC‐exshCtr). All data are presented as means ± SEM, Mann‐Whitney test. PBS, phosphate saline solution; MSC‐ex, mesenchymal stem cells derived exosomes
FIGURE 4
FIGURE 4
Ang‐2 enhanced HDLEC lymphangiogenesis by upregulating Prox1 mediated Akt signaling. (A) Western blot for Ang2 in HDLEC transfected with lenti‐GFP or lenti‐Ang2 (Ai), control shRNA (shCtr) or Ang2‐shRNA (shAng2) (Aii). (B, C) Transwell migration assay, matrigel tube formation of HDLEC transfected with lenti‐GFP or lenti‐Ang2. (n = 4; *P < .05 vs GFP, **P < .01 vs GFP). (D, E) Transwell migration assay, matrigel tube formation of HDLEC transfected with shCtr or shAng2. (n = 4; *P < .05 vs shCtr, ns, not significant). (F, G) Cell proliferation of HDLEC transfected with lenti‐GFP or lenti‐Ang2 (F), shCtr or shAng2 (G) by CCK8 assay. (n = 4; *P < .05 vs GFP, **P < .01 vs GFP). (H) Western blot for Prox1, p‐Akt, and t‐Akt in HDLEC transfected with lenti‐GFP or lenti‐Ang2 (n = 4; *P < .05 vs GFP). (I, J) Western blot for Prox1, p‐Akt in HDLEC transfected with lenti‐GFP, lenti‐Prox1, control shRNA (shCtr), Prox1‐shRNA (shProx1) (n = 4; *P < .05 vs GFP). (K) Western blot for p‐Akt in HDLEC transfected with lenti‐GFP, lenti‐Prox1, lenti‐Prox1/MK‐2206 (Akt inhibitor) treatment (n = 4; *P < .05 vs lenti‐Prox1). (L) Matrigel tube formation of HDLEC transfected with lenti‐GFP, lenti‐Prox1, lenti‐Prox1/MK‐2206 treatmen,t and quantification (n = 4; *P < .05 vs GFP, # P < .05 vs Prox1). Scale bars, 50 μm. All data are presented as means ± SEM, Mann‐Whitney test. (M) Schematic representation of hypothetic mechanism for Ang2‐enriched MSC‐ex mediating prolymphangiogenesis through upregulating Prox1 mediated Akt signaling
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References

    1. Aldrich MB, Rasmussen JC, Fife CE, Shaitelman SF, Sevick‐Muraca EM. The development and treatment of lymphatic dysfunction in cancer patients and survivors. Cancers. 2020;12:2280. - PMC - PubMed
    1. The global distribution of lymphatic filariasis, 2000–18: a geospatial analysis. Lancet Glob Health. 2020;8(9):e1186‐e1194. - PMC - PubMed
    1. Schaverien MV, Aldrich MB. New and emerging treatments for lymphedema. Semin Plast Surg. 2018;32(1):48‐52. - PMC - PubMed
    1. Rutkowski JM, Boardman KC, Swartz MA. Characterization of lymphangiogenesis in a model of adult skin regeneration. Am J Physiol Heart Circ Physiol. 2006;291(3):H1402‐10. - PMC - PubMed
    1. Jiang X, Nicolls MR, Tian W, Rockson SG. Lymphatic dysfunction, leukotrienes, and lymphedema. Annu Rev Physiol. 2018;80:49‐70. - PMC - PubMed

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