Indication-based protocols with different solutions for mechanical stromal-cell transfer

Abstract

Background Regenerative medicine is the fastest developing branch of plastic surgery in recent times. Adipose tissue is one of the largest and most important sources in the body for stromal cells. Although mechanical isolation methods are both very popular and have many advantages, they still have no accepted protocols. Objective We developed new protocols called indication-based protocols (IPs) for standardization and new techniques called mechanical stromal-cell transfer (MEST) by using ultra-sharp blades and dilution of adipose tissue with different solutions (saline, Ringer and 5% Dextrose) Methods & material: In order to obtain the desired physical structure (liquid, gel, solid) and the desired volume, four different types of IPs have been defined. Adipose tissue was prediluted with different solutions using 10 or 20 cc injectors in IPs 1 and 2, while condensed adipose tissue was used directly in IPs 3 and 4. Results In MEST, stromal cells were obtained from 100 mL of condensed fat using different IPs with 92% mean viability and cell counts of 26.80–91.90 × 106. Stromal cells can be obtained in the desired form and number of cells by using four different IPs. Conclusion Isolation of stromal cells by cutting fat with sharp blades will prevent the death of fat tissue and stromal cells and will allow high viability and cell count with our new technique. Predilution with different solutions: Diluting the condensed adipose tissue with the desired solutions (saline, Ringer or 5% Dextrose) before the adinizing process will provide even more stromal cells. Lay Summary Obtaining regenerative stromal cells from adipose tissue can be done by two methods: Enzymatic and mechanical. Mechanical methods have many advantages. Although mechanical stromal cell extraction from adipose tissue is very popular and many techniques have been described, there are still no accepted protocols, definition for the end product, and no consensus on the status of the stromal cells. In this study, stromal cells were obtained mechanically by using ultra-sharp blade systems, without exposing adipose tissue to blunt trauma. Thus, a higher number of cells and higher viability could be obtained. An “Indication based” protocol has been defined for the first time in order to obtain the desired number and status (solid, semi-solid, liquid) end product. Diluting the condensed adipose tissue with the desired solutions (saline, Ringer or 5% Dextrose) before the adinizing process will provide even more stromal cells. This will provide an opportunity for clinicians to obtain and apply a stromal cell solution for different indications in different anatomical regions.

In Vivo Evaluation of Mechanically Processed Stromal Vascular Fraction in a Chamber Vascularized by an Arteriovenous Shunt

“…These methods circumvent regulatory restraints by utilizing mechanical forces without the employment of enzymes and skipping the lengthy cultivation process [6]. Several mechanical protocols were suggested which include shaking [7], extended centrifugation [8], vibration [9], cutting with blades [10], emulsification [11] and many more. In 2019, the International Federation for Adipose Therapeutics and Science (IFATS) stressed the particular importance of mechanically processed SVF [12].…”

A New Classification for Adipose-derived Stromal-cell Systems

“…11 This approach will be highly advantageous in male and female genital applications, where a 400/600 μm diameter fat graft will be applied in thin-skinned and superficial areas, and at the same time significant skin improvement will be achieved with regeneration. 14 Similarly, while large-diameter fat grafting can be placed intramuscularly in breast tissue, applying a smaller diameter fat graft in areas closer to the surface will both increase the chance of success and reduce the risk of complications such as fat necrosis and microcalcification. 15 This approach applies to high-volume gluteal fat grafting.…”

Comparison of the Effectiveness of Methods for Obtaining and Application the Stromal-Vascular Fraction of Adipose Tissue in an Experimental Model of a Donor Wound in Burns

“…Ферментативное расщепление при помощи коллагеназы остается «золотым стандартом», поскольку обеспечивает получение большего количества ММСК из единицы объема жировой ткани [6][7][8]. Механическая обработка требует существенно меньших материальных и временных затрат, однако технологии получения СВФ таким способом трудно стандартизировать [9][10][11].…”