Park In-Su
Cell Therapy Center, Ajou University School of Medicine, Suwon, Korea.
Int J Stem Cells. 2021 May 30;14(2):212-220. doi: 10.15283/ijsc20175.
The objective of this study was to investigate whether conditioned medium from photobiomodulation (PBM) irradiated adipose-derived stromal cell (ASC) spheroids prior to implanting could stimulate angiogenesis and tissue regeneration to improve functional recovery of skin tissue in an animal skin wound model.
ASC were split and seeded on chitosan-coated 24 well plate at a density of 7.5×10 cells/cm, and allowed to adhere at 37°C. Within 3 days of culture, ASC formed spheroids by PBM irradiation. Conditioned medium (CM) fractions were collected from the PBM-ASC to yield nor adipose-derived stromal cell spheroid (spheroid) and PBM-spheroid, respectively, centrifuged at 13,000 g at 4℃ for 10 min, and stored prior to use for ELISA, protein assay, or in vivo wound-healing assays. Phosphate-buffered saline, cultured CM from ASCs, PBM irradiation prior to implanting conditioned medium from ASC, cultured CM from ASC spheroid, and PBM-spheroid-CM (PSC) were transplanted into a wound bed in athymic mice to evaluate therapeutic effects of PSC . PSC enhanced wound closure in a skin injury model compared to PBS, CM, PBM-CM, and spheroid-CM. The density of vascular formations increased as a result of angiogenic factors released by the wound bed and enhanced tissue regeneration at the lesion site.
These results indicate that implant of PSC can significantly improve functional recovery compared to PBS, CM, PBM-CM, or spheroid-CM treatment. Implant of PSC may be an effective form of paracrine mediated therapy for treating a wound bed.
本研究的目的是调查在植入前,光生物调节(PBM)照射的脂肪来源间充质干细胞(ASC)球体的条件培养基是否能刺激血管生成和组织再生,以改善动物皮肤伤口模型中皮肤组织的功能恢复。
将ASC消化并以7.5×10个细胞/cm的密度接种到壳聚糖包被的24孔板上,在37℃下使其贴壁。在培养的3天内,通过PBM照射使ASC形成球体。分别从PBM-ASC收集条件培养基(CM)组分,得到非脂肪来源间充质干细胞球体(球体)和PBM-球体,在4℃下以13,000 g离心10分钟,储存备用,用于ELISA、蛋白质测定或体内伤口愈合试验。将磷酸盐缓冲盐水、ASC培养的CM、植入ASC条件培养基前的PBM照射、ASC球体培养的CM和PBM-球体-CM(PSC)移植到无胸腺小鼠的伤口床中,以评估PSC的治疗效果。与PBS、CM、PBM-CM和球体-CM相比,PSC在皮肤损伤模型中增强了伤口闭合。由于伤口床释放的血管生成因子,血管形成密度增加,病变部位的组织再生增强。
这些结果表明,与PBS、CM、PBM-CM或球体-CM治疗相比,植入PSC可显著改善功能恢复。植入PSC可能是一种有效的旁分泌介导疗法,用于治疗伤口床。