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

Independent University studies have validated Bright R. et al. patented sonication protocol to separate mesenchymal stem cells from adipose tissue. The obtain cells have proven to be

  • Viable
  • Adherent
  • Capable of mitotic division and
  • Differentiation into multiple cell types

There were no differences between collagenase digestion and Bright R. et al. 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

We have differentiated the stem cells into 3 different tissue types to prove their ability. These are listed below.

  1. Cartilage
  2. Bone
  3. Fat

1. 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

2. 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

3. 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

 

 

Further information

To obtain further information about the stem cell treatment and Macquarie Stem Cells, please click on the link below.

Supporting Information