Isolation and Maintenance of Adipose Stem Cell

Adipose tissue is a rich source of mesenchymal stem cells (MSC). Given their easy accessibility and abundant source, adipose stem cells (ASCs) have demonstrated great potential in the stem cell field. Although many scientists have successfully isolated ASCs, there is no standardized protocol for ASC isolation, maintenance, and culture.

Creative Biolabs describes a protocol for the isolation and maintenance of ASCs. The purpose of this protocol is to isolate and proliferate stem cells from adipose tissue. This is a relatively simple procedure that we consider in terms of quality, safety, and efficacy.

Materials Required

• Blood/saline solution containing adipose tissue
• Collagenase (type I, IA, II or IV)
• Basic culture medium
• 1X phospho-buffered saline
• Sterile glassware and plastic utensils

Procedure

Washing

Wash the harvested fat aspirate with sterile PBS until golden yellow in color.

Digestion

Add the collagenase solution to the fat aspirate proportionally for digestion. Incubate on a shaker for several hours until it becomes cloudy. After enzyme treatment, the floating adipocytes, lipids and fluids can be aspirated, leaving the stromal vascular fraction (SVF) particles.

Isolation

Collect SVF by centrifugation and resuspend SVF precipitate for a few minutes of incubation at room temperature. Then, centrifuge and collect the cell precipitate again. Next, resuspend the cell precipitate and filter out the larger tissue particles through a sieve. Finally, centrifuge to obtain the cell precipitate.

Cultivation

Resuspend the SVF pellet containing ASC and dispense the suspension by pipetting into the tissue culture treated dishes. Incubate cells at 37°C, 5% CO₂ for 3-4 days without changing medium.

Notes

• The type of collagenase used varies from experiment to experiment, with type IA collagenase being more common.
• Digestion time for cell isolation from adipose tissue varies from study to study. We recommend a typical digestion time of 1-2 hours.
• Filtration may be necessary due to the presence of large fibrillated material and cell aggregates after digestion.
• Some steps can be taken to remove as many erythrocytes as possible.

We describe our protocols in a simple and clear manner for our clients to follow. You can also get stem cell isolation services at a low cost by contacting us directly.

References

1. Zhu M, et al. Manual isolation of adipose-derived stem cells from human lipoaspirates. J Vis Exp, 2013; (79): e50585.
2. Helmy M A, et al. A protocol for primary isolation and culture of adipose-derived stem cells and their phenotypic profile. Alexandria Journal of Medicine, 2020, 56(1), 42-50.

For Research Use Only. Not For Clinical Use.