Muscle progenitor stem cells in 3D maintain character after expansion in Mimetix scaffolds

CD133+ cell growth and distribution at D7 (A) and D14 (C) in 2D culture and in 3D scaffold at D7 (B) and D14 (D). The images of the
CD133+ cells in 2D culture or in 3D scaffolds were obtained by inverted light fluorescence microscopy using a Leica DMI6000B microscope: panel 1:
bright field (BF); panel 2: DAPI staining (blue); panel 3: calcein staining (green). Arrowheads in (C) highlight the myotube formation.scaffold at D7 (B) and D14 (D). The images of the
CD133+ cells in 2D culture or in 3D scaffolds were obtained by inverted light fluorescence microscopy using a Leica DMI6000B microscope: panel 1:
bright field (BF); panel 2: DAPI staining (blue); panel 3: calcein staining (green). Arrowheads in (C) highlight the myotube formation. Courtesy L Al-Khalili, KTH.

CD133+ progenitor stem cells were shown to maintain ‘stem-ness’ after expansion in Mimetix 3D cell culture.

In a recent publication in the Journal of Biomaterials and Tissue Engineering, researchers at KTH in Sweden characterised the growth and differentiation of CD133+ stem cells in Mimetix 3D and in conventional 2D cell culture.

CD133+ cells are myogenic progenitors for skeletal muscle regeneration. If they can be expanded in culture while maintaining their stem cell character they have the potential to be used to treat muscular dystrophies.

In the study the proliferation of human CD133+ cells was studied for 14 days in Mimetix scaffolds (3D biomimetic electrospun poly-L-lactic acid (PLLA) nano-fiber scaffolds). Additionally, the myogenic differentiation of the cells was studied during the last 7 days of the culture period. The cells were homogeneously distributed in the 3D scaffolds while colony formation and myotube formation occurred in 2D. The marker expression during proliferation and differentiation, together with the absence of myotubes in 3D, indicates a better maintenance of stemness in 3D PLLA and stronger tendency for spontaneous differentiation in 2D culture.

One of the greatest challenges for regenerative medicine therapies is to obtain large cell numbers of adequate quality. It appears that the electrospun scaffold is a promising avenue to explore for large scale expansion of CD133+ cells given the advantages of significant cell expansion capacity and the resistance to spontaneous and induced differentiation.

The research leading to these results was supported by the European Union’s seventh framework programma FP7/Health 2013 Innovation 2 under grant agreement 601700 HESUB.

Reference: Characterisation of Human CD133+ Cells in Biocompatible Poly(l-lactic acid) Electrospun Nano-fiber scaffolds. Lubna Al-Khalili, Karin Gillner, Ye Zhang, Carolina Åstrand, Atefeh Shokri,
Nanaaya Hughes-Brittain, Robert McKean, Brendan Robb, and Veronique Chotteau. Journal of Biomaterials and Tissue Engineering Vol. 6, 959-966, 2016.

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