Rapid and effective preparation of clonal bone marrow-derived mesenchymal stem/stromal cell sheets to reduce renal fibrosis

• Published in: Scientific reports Vol. 13; no. 1; p. 4421
• Main Authors: Kameishi, Sumako, Dunn, Celia M., Oka, Masatoshi, Kim, Kyungsook, Cho, Yun-Kyoung, Song, Sun U., Grainger, David W., Okano, Teruo
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 17.03.2023; Nature Publishing Group; Nature Portfolio
• Subjects: 631/532/2074; 631/61/2035; 631/61/2320; Animals; Bone Marrow; Cell culture; Cell suspensions; Fibrosis; Freeze-thawing; Gene expression; Growth factors; Hepatocyte growth factor; Humanities and Social Sciences; Humans; Interleukin 10; Ischemia; Kidney diseases; Kidney Diseases - metabolism; Kidney Diseases - therapy; Medical treatment; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; multidisciplinary; Rats; Reperfusion; Science; Science (multidisciplinary); Thawing; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - metabolism
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-023-31437-7

Allogeneic “off-the-shelf” mesenchymal stem/stromal cell (MSC) therapy requires scalable, quality-controlled cell manufacturing and distribution systems to provide clinical-grade products using cryogenic cell banking. However, previous studies report impaired cell function associated with administering freeze-thawed MSCs as single cell suspensions, potentially compromising reliable therapeutic efficacy. Using long-term culture-adapted clinical-grade clonal human bone marrow MSCs (cBMSCs) in this study, we engineered cBMSC sheets in 24 h to provide rapid preparation. We then sought to determine the influence of cBMSC freeze-thawing on both in vitro production of pro-regenerative factors and in vivo ability to reduce renal fibrosis in a rat model compared to freshly harvested cBMSCs. Sheets from freeze-thawed cBMSCs sheets exhibited comparable in vitro protein production and gene expression of pro-regenerative factors [e.g., hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), and interleukin 10 (IL-10)] to freshly harvested cBMSC sheets. Additionally, freeze-thawed cBMSC sheets successfully suppressed renal fibrosis in vivo in an established rat ischemia–reperfusion injury model. Despite previous studies reporting that freeze-thawed MSCs exhibit impaired cell functions compared to fresh MSC single cell suspensions, cell sheets engineered from freeze-thawed cBMSCs do not exhibit impaired cell functions, supporting critical steps toward future clinical translation of cBMSC-based kidney disease treatment.