Jacobs Journal of Bone Marrow and Stem Cell Research

An Efficient Method for Rapid Establishment of Fibroblast-Like Cells from Horse Subcutaneous Adipose Tissue

Published on: 2016-03-04

Abstract

Adipose-tissue derived stromal cells (ADSCs) are easily obtainable multipotent mesenchymal stem cells capable of differentiation into various cell lineages making it an attractive cell source for regenerative medicine. ADSC is generally isolated from subcutaneous adipose tissue by enzymatic digestion of biopsies followed by removal of undigested tissues and seeding of the dissociated cells onto plastic. This is an inefficient procedure where only a tiny fraction of cells are established in culture and rest of the cells are lost. Development of new methods would be desirable to recover most cells of the originally collected biopsies. This would reduce the time between biopsy collection and transplantation by producing large quantities of cells within a short time span. One of the benefits of this method would be to provide autologous cells for quick replacement therapies. Present report describes a new cell culture method which allows establishment of almost all cells of adipose tissue biopsies on plastics and the adherent cells show fibroblast-like morphology with rapid self-renewal capacity. ADSCs of subcutaneous fat tissue of horse can be isolated efficiently by mincing the excised biopsy with a surgical scissor and culturing of fragmented tissue in RPMI 1640 medium containing 15% ordinary fetal bovine serum for 2-3 weeks without change of medium. The conditioned media of highly proliferating ADSCs not only maintain self-renewal but also improve proliferation of other cells indicating that proliferating ADSCs secrete growth promoting factors in the culture media. The new method has direct application both in basic research and in regenerative medicine. The new method is named as “conditioned media-mediated Cell Amplification, (cmCellAmp)”.

Keywords

Adipose-Tissue Derived Stromal Cell; Adult Stem Cell; Embryonic Stem Cell; Autologous Transplantation; Immortality; Conditioned Media; Fibroblast; Cell Multiplication