Isolation of human foetal myoblasts and its application for microencapsulation

Abstract

Foetal cells secrete more growth factors, generate less immune response, grow and proliferate better than adult cells. These characteristics make them desirable for recombinant modification and use in microencapsulated cellular gene therapeutics. We have established a system in vitro to obtain a pure population of primary human foetal myoblasts under several rounds of selection with non-collagen coated plates and identified by desmin staining. These primary myoblasts presented good proliferation ability and better differentiation characteristics in monolayer and after microencapsulation compared to murine myoblast C2C12 cells based on creatine phosphokinase (CPK), major histocompatibility complex (MHC) and multi-nucleated myotubule determination. The lifespan of primary myoblasts was 70 population doublings before entering into senescent state, with a population time of 18–24 hrs. Hence, we have developed a protocol for isolating human foetal primary myoblasts with excellent differentiation potential and robust growth and longevity. They should be useful for cell-based therapy in human clinical applications with microencapsulation technology.

Potentiated Osteoinductivity via Cotransfection with BMP-2 and VEGF Genes in Microencapsulated C2C12 Cells

“…The C2C12 cell line acts as a premyoblast cell line [ 26 ] and could be shifted towards osteoblast differentiation from myoblast differentiation with interference of the BMP-2 protein [ 27 – 29 ]. Moreover, Li et al [ 30 ] proposed that C2C12 cells were able to secrete cytokines and survive within incapacious microcapsules in the long term. Therefore, further research was carried out to compare the osteoinductivity of C2C12 cells with a single gene transfection with those with a double gene cotransfection.…”

3D Printed porous polyamide macrocapsule combined with alginate microcapsules for safer cell-based therapies

“…We produced APA microcapsules containing either C 2 C 12 -EPO myoblasts or BHK-VEGF fibroblasts. These cell lines have successfully been used in preclinical models using different encapsulation systems for the secretion of recombinant proteins 40 – 43 . Also, in our experience, these two types of cells grow without problems inside alginate scaffolds and can be used either for in vitro or in vivo experiments without losing their functionality 44 .…”