Molecular characterization of hematopoietic stem cells after in vitro amplification on biomimetic 3D PDMS cell culture scaffolds

• Published in: Scientific reports Vol. 11; no. 1; p. 21163
• Main Authors: Marx-Blümel, Lisa, Marx, Christian, Sonnemann, Jürgen, Weise, Frank, Hampl, Jörg, Frey, Jessica, Rothenburger, Linda, Cirri, Emilio, Rahnis, Norman, Koch, Philipp, Groth, Marco, Schober, Andreas, Wang, Zhao-Qi, Beck, James F.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 27.10.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 1-Phosphatidylinositol 3-kinase; 631/337/2019; 631/45/475; 631/532/1542; 631/80/86; 639/301/357/537; Adaptation; Adult; AKT protein; Biomimetic Materials - adverse effects; Biomimetic Materials - chemistry; Biomimetics; Bone marrow; Cell culture; Cell Proliferation; Cells, Cultured; Dimethylpolysiloxanes - adverse effects; Dimethylpolysiloxanes - chemistry; Female; Forkhead protein; Forkhead Transcription Factors - metabolism; Hematopoietic stem cells; Hematopoietic Stem Cells - drug effects; Hematopoietic Stem Cells - metabolism; Hematopoietic Stem Cells - physiology; Humanities and Social Sciences; Humans; Hypoxia-Inducible Factor 1, alpha Subunit - metabolism; Male; Metabolic flux; Metabolism; multidisciplinary; Phosphatidylinositol 3-Kinases - metabolism; Pluripotency; Polydimethylsiloxane; Proteomes; Proto-Oncogene Proteins c-akt - metabolism; Science; Science (multidisciplinary); Signal Transduction; Stem cells; Sterol Regulatory Element Binding Proteins - metabolism; Sterol regulatory element-binding protein; Tissue Scaffolds - adverse effects; Tissue Scaffolds - chemistry; TOR protein; TOR Serine-Threonine Kinases - metabolism; Transcriptome; Transcriptomes; Transplantation
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-021-00619-6

Hematopoietic stem cell (HSC) transplantation is successfully applied since the late 1950s. However, its efficacy can be impaired by insufficient numbers of donor HSCs. A promising strategy to overcome this hurdle is the use of an advanced ex vivo culture system that supports the proliferation and, at the same time, maintains the pluripotency of HSCs. Therefore, we have developed artificial 3D bone marrow-like scaffolds made of polydimethylsiloxane (PDMS) that model the natural HSC niche in vitro. These 3D PDMS scaffolds in combination with an optimized HSC culture medium allow the amplification of high numbers of undifferentiated HSCs. After 14 days in vitro cell culture, we performed transcriptome and proteome analysis. Ingenuity pathway analysis indicated that the 3D PDMS cell culture scaffolds altered PI3K/AKT/mTOR pathways and activated SREBP, HIF1α and FOXO signaling, leading to metabolic adaptations, as judged by ELISA, Western blot and metabolic flux analysis. These molecular signaling pathways can promote the expansion of HSCs and are involved in the maintenance of their pluripotency. Thus, we have shown that the 3D PDMS scaffolds activate key molecular signaling pathways to amplify the numbers of undifferentiated HSCs ex vivo effectively.