Nonviral Cell Labeling and Differentiation Agent for Induced Pluripotent Stem Cells Based on Mesoporous Silica Nanoparticles

Abstract

The generation of induced pluripotent stem cells (iPSCs) is an innovative personalized-regenerative technology, which can transform own-self somatic cells into embryonic stem (ES)-like cells, which have the potential to differentiate into all cell types of three dermal lineages. However, how to quickly, efficiently, and safely produce specific-lineage differentiation from pluripotent-state cells and iPSCs is still an open question. The objective of the present study was to develop a platform of a nonviral gene delivery system of mesoporous silica nanoparticles (MSNs) to rapidly generate iPSC-derived definitive-lineage cells, including endodermal-differentiated cells. We also evaluated the feasibility and efficiency of FITC-conjugated MSNs (FMSNs) for labeling of iPSCs and utilized the multifunctional properties of FMSNs for a suitable carrier for biomolecule delivery. We showed that FMSNs of various surface charges could be efficiently internalized by iPSCs without causing cytotoxicity. The levels of reactive oxygen species and pluripotent status, including in vitro stemness signatures and in vivo teratoma formation, remained unaltered. Notably, positive-charged FMSN enhanced cellular uptake efficiency and retention time. Moreover, when using positive-charged FMSN to deliver hepatocyte nuclear factor 3β (HNF3β) plasmid DNA (pDNA), the treated iPSCs exhibited significantly improved definitive endoderm formation and further quickly differentiated into hepatocyte-like cells with mature functions (low-density lipoprotein uptake and glycogen storage) within 2 weeks in vitro. Double delivery of pHNF3β further improved mRNA expression levels of liver-specific genes. These findings reveal the multiple advantages of FMSNs to serve as ideal vectors not only for stem cell labeling but also for safe gene delivery to promote the production of hepatocyte-like cells from iPSCs.

A biomaterial approach to cell reprogramming and differentiation

“…Furthermore, MSNs are known to have a highly porous internal structure, which provides large surface area for loading high amounts of molecular cargo within the nanoparticle. MSNs were advantageously used for the rapid differentiation of miPSCs to definitive lineage cells (Figure 4D) [37]. To be specific, non-viral vectors encoding hepatocyte nuclear factor 3β (pHNF3β) were electrostatically adsorbed to positively charged FITC-labeled MSNs (FMSNs).…”

Facile Synthesis of Yolk-Shell-Structured Triple-Hybridized Periodic Mesoporous Organosilica Nanoparticles for Biomedicine

“…Mesoporous silica with the properties of high surface area, large pore volume, uniform and tunable pore size, and easy functionalization has attracted great attention in biomedical applications such as drug/gene/protein delivery, [1][2][3][4][5][6][7][8][9] diagnostic imaging, [10] theranostics, [11] and functional implants. [12][13][14] The enhanced therapeutic efficacy, reduced side effects, little biotoxicity, and degradation of mesoporous silica in the biomedical applications have been preliminary demonstrated.…”

Vectorized nanodelivery systems for ischemic stroke: a concept and a need

“…Likewise, nanomedicine may be viewed as a necessary complement of cell-based therapies due to excellent biocompatibility, ease of cellular internalization, and in vivo cell tracking possibilities. Nanomedicine may be a potential drug delivery platform used to induce specific differentiation- and cell-oriented therapeutics [148, 151–154]. …”