A New Nonenzymatic Method and Device to Obtain a Fat Tissue Derivative Highly Enriched in Pericyte-Like Elements by Mild Mechanical Forces from Human Lipoaspirates

Abstract

Adipose tissue contains multipotent elements with phenotypic and gene expression profiles similar to human mesenchymal stem cells (hMSCs) and pericytes. The chance of clinical translation of the multilineage potential of these cells is delayed by the poor/negligible cell survival within cryopreserved lipoaspirates, the difficulty of ex vivo expansion, and the complexity of current Good Manufacturing Practice (cGMP) requirements for expanded cells. Hence, availability of a minimally manipulated, autologous, hMSC/pericyte-enriched fat product would have remarkable biomedical and clinical relevance. Here, we present an innovative system, named Lipogems, providing a nonexpanded, ready-to-use fat product. The system uses mild mechanical forces in a completely closed system, avoiding enzymes, additives, and other manipulations. Differently from unprocessed lipoaspirate, the nonexpanded Lipogems product encompasses a remarkably preserved vascular stroma with slit-like capillaries wedged between adipocytes and stromal stalks containing vascular channels with evident lumina. Immunohistochemistry revealed that Lipogems stromal vascular tissue included abundant cells with pericyte/hMSC identity. Flow cytometry analysis of nonexpanded, collagenase-treated Lipogems product showed that it was comprised with a significantly higher percentage of mature pericytes and hMSCs, and lower amount of hematopoietic elements, than enzymatically digested lipoaspirates. Differently from the lipoaspirate, the distinctive traits of freshly isolated Lipogems product were not altered by cryopreservation. Noteworthy, the features of fresh product were retained in the Lipogems product obtained from human cadavers, paving the way to an off-the-shelf strategy for reconstructive procedures and regenerative medicine. When placed in tissue culture medium, the Lipogems product yielded a highly homogeneous adipose tissue-derived hMSC population, exhibiting features of hMSCs isolated from other sources, including the classical commitment to osteogenic, chondrogenic, and adipogenic lineages. Moreover, the transcription of vasculogenic genes in Lipogems-derived adipose tissue hMSCs was enhanced at a significantly greater extent by a mixture of natural provasculogenic molecules, when compared to hMSCs isolated from enzymatically digested lipoaspirates.

Non-Responding Knee Pain with Osteoarthritis, Meniscus and Ligament Tears Treated with Ultrasound Guided Autologous, Micro-Fragmented and Minimally Manipulated Adipose Tissue

“…The concept that fat tissue is an optimal source for MSCs which is now established, supported by their abundance and compared to other tissues [11] [12] [13]. Indeed, 1/100 cells in the adipose tissue are MSCs compared to 1/100,000 cells in the bone marrow.…”

“…In less manipulated fat tissue are preserved both cells and the niche: a natural adipose scaffold which is the "perfect functional unit" [11].…”

“…The fat tissue is treated mechanically without expansion and/or enzymatic treatment. Lipogems® device provides autologous, micro-fragmented, and minimally manipulated adipose tissue graft [11] [21].…”

Adipose Tissue-Derived Mesenchymal Stem Cells: A Novel Therapy in Orthopedics

“…Researchers investigated and tested clinically the disadvantages of the bone marrow derived mesenchymal stem cells (hBMSCs) by enzymatic lipoaspiration process [40]. The aforementioned procedure is painful and invasive [40].…”

“…The aforementioned procedure is painful and invasive [40]. Its cell population yield is low and the cell fate in vivo is uncertain, especially that the delivery efficiency is reduced [20,[40][41][42][43][44][45][46].…”