18.2.6 - Multiple Nanosecond Pulsed Electric Fields Enhanced Multilineage Differentiation Potential of MSCs during Prolonged In Vitro Culture
- Z. Ge (Beijing, CN)
- Z. Ge (Beijing, CN)
- J. Chen (Beijing, CN)
Abstract
Purpose
Purpose：Mesenchymal stem cells (MSCs) gradually lose multilineage differentiation potentials when cultured in vitro, which hinders their subsequent applications. Though much efforts have been enforced to take this challenge, progresses are limited. In our previous research, nanosecond pulsed electrical fields (nsPEFs）enhanced multilineage differentiation potential of MSCs. We tried to explore if concomitant applications of nsPEFs could enhanced differentiation potentials of MSCs during prolonged in vitro culture.
Methods and Materials
Materials & Methods: Pig mesenchymal stem cells cultured on conductive films were treated with nanosecond pulsed electrical fields (nsPEFs). After optimizing parameters (100ns 10kv/cm, 10ns 20kv/cm, judged by gene expression of Oct4A, Nanog and Sox2) of nsPEFs on conductive film, concomitant applications of multiple nsPEFs treatment (four times) through conductive film were performed.
Results
Results: In this study, pre-conditioning of mesenchymal stem cells with nanosecond pulsed electrical fields (nsPEFs) through gas cuvettes significantly enhanced the expression of Oct4A (3.8 fold of 100ns 10kv/cm, 3.4 fold of 10ns 20kv/cm), Nanog (3.2 fold of 100ns 10kv/cm, 2.8 fold of 10ns 20kv/cm) and Sox2 (1.8 fold of 100ns 10kv/cm, 2.0 fold of 10ns 20kv/cm). Furthermore, after optimizing parameters of nsPEFs on conductive film (100ns 10kv/cm) and time interval (3d between each nsPEFs treatment), concomitant applications of multiple nsPEFs treatment (four times) enhanced gene expression of Oct4A (3.5~4.5fold), Nanog (3.5~4fold) and Sox2 (1.3~2fold) significantly while maintained this high level gene expression for nearly 30 days of MSCs, much longer than single nsPEFs treatment (normal 7 days). Differentiation potential was significantly increased in both single and multiple nsPEFs treatment, judged by chondrogenic (3-fold of Sox9), osteogenic (6-fold Runx2) and adipogenic (4-fold of PPAR-γ) marker gene.
Conclusion
Conclusion: Our study suggested that multiple nsPEFs treatment through conductive film could enhance the multilineage differentiation potential of MSC for long time scale.