Independent University Studies on Stem cell Viability (live cells) and Differentiation on SVF Separation by Australian Patented Sonication Protocol

Independent University studies have validated Cell-Innovations sonication protocol to obtain viable, adherent cells capable of mitotic division and differentiation indicative of mesenchymal stem cells.

There were no differences between collagenase digestion and Cell-Innovations sonication protocol in terms of cell population doubling time, freeze-thaw viability, or differentiation into either bone or adipose tissues.

The old methods of collagenase separation of adipose to SVF are shown below in comparison to sonication separation of SVF in the University study below.

Cells Viable and Cultured From Adipose SVF

Adipose derived SVF after 5 days in culture at 40x magnification. Images obtained of cells isolated from patients using collagenase correspond to: a, c, and e, respectively. Images obtained of cells isolated from patients using sonication correspond to: b, d, and f, respectively.

adipose

Differentiation Slide Images

Osteoblastic (bone) differentiation of stem cells are typically tested by staining with Alizarin red which indicates calcium-rich deposits.

Adipose derived SVF cells cultured in osteogenic media for 28 days (bii-eii) or control media for 6 days (bi-ei) from patients and stained with alizarin red. The stain alizarin red Images at 100x magnification. Images b, and d were obtained from cells isolated using collagenase method. Images c, and e, were obtained from cells isolated using sonication.

bone

Adipogenic (fat cell) differentiation of mesenchymal stem cells are typically tested by staining with Oil Red O which stains triglycerides (intracellular lipid vesicles) indicative of adipocyte (fat) cells.

Adipose derived SVF cells cultured in adipogenic media for 12 days (bii-eii) or control media for 6 days (bi-ei) from patients and stained with oil red o. Images at 100x magnification. Images b, and d were obtained from cells isolated using collagenase method. Images c, and e, were obtained from cells isolated using sonication.

adipogenic

Chondrogenic (cartilage) differentiation of mesenchymal stem cells results in formation of cartilage with a typical extracellular matrix. A key molecule besides collagen type II within this extracellular matrix is the proteoglycan aggrecan. Alcian Blue a blue copper containing dye can detect aggrecan an indicator of cartilage formation.

culture

More Information

Stem Cell Outcomes
Clinical Trials