Characterization of freshly isolated and cultured cells derived from the fatty and fluid portions of liposuction aspirates

Abstract

Liposuction aspirates (primarily saline solution, blood, and adipose tissue fragments) separate into fatty and fluid portions. Cells isolated from the fatty portion are termed processed lipoaspirate (PLA) cells and contain adipose-derived adherent stromal cells (ASCs). Here we define cells isolated from the fluid portion of liposuction aspirates as liposuction aspirate fluid (LAF) cells. Stromal vascular fractions (SVF) were isolated separately from both portions and characterized under cultured and non-cultured conditions. A comparable number of LAF and PLA cells were freshly isolated, but fewer LAF cells were adherent. CD34+ CD45- cells from fresh LAF isolates were expanded by adherent culture, suggesting that LAF cells contain ASCs. Although freshly isolated PLA and LAF cells have distinct cell surface marker profiles, adherent PLA and LAF cells have quite similar characteristics with regard to growth kinetics, morphology, capacity for differentiation, and surface marker profiles. After plating, both PLA and LAF cells showed significant increased expression of CD29, CD44, CD49d, CD73, CD90, CD105, and CD151 and decreased expression of CD31 and CD45. Multicolor FACS analysis revealed that SVF are composed of heterogeneous cell populations including blood-derived cells (CD45+), ASCs (CD31- CD34+ CD45- CD90+ CD105- CD146-), endothelial (progenitor) cells (CD31+ CD34+ CD45- CD90+ CD105low CD146+), pericytes (CD31- CD34- CD45- CD90+ CD105- CD146+), and other cells. After plating, ASCs showed a dramatic increase in CD105 expression. Although some adherent ASCs lost CD34 expression with increasing culture time, our culture method maintained CD34 expression in ASCs for at least 10-20 weeks. These results suggest that liposuction-derived cells may be useful and valuable for cell-based therapies.

Concise Review: Adipose-Derived Stromal Vascular Fraction Cells and Stem Cells: Let's Not Get Lost in Translation

“…The development of an efficient and reproducible mechanical-based tissue disruption process would remove the need for enzyme reagents and merits further investigation. There is evidence that functional ASCs can be expanded directly from lipoaspirate fluids without the need for collagenase digestion [14]. Similarly, multiple groups routinely use porcine-derived trypsin to passage plastic adherent ASCs, and recent studies have documented the equivalent performance of bacterial-derived or corn-derived trypsin products [15][16][17].…”

Characterization of Human Adipose Tissue-Resident Hematopoietic Cell Populations Reveals a Novel Macrophage Subpopulation with CD34 Expression and Mesenchymal Multipotency

“…The SVF was isolated from the aspirated AT as described previously [25]. Briefly, the aspirated AT was washed with phosphate-buffered saline (PBS) and digested on a shaker at 37°C in PBS containing 0.075% collagenase (crude type, cat No.…”

“…The methods of differentiation and analyses have been confirmed to be reliable for induction of ASC differentiation [25]. Cells cultured in the control medium [the Dulbecco's modified Eagle's medium (DMEM) containing 10% FBS] were used as negative controls.…”

Phenotypes of stem cells from diverse origin

“…Therefore, we asked all authors to revisit the table and upgrade their part either by own data or based on additional literature, as necessary. This resulted in the upgraded Table 1 showing phenotypes of very small embryonic-like stem cells (vSELs) (5,6), neural stem cells (NSCs) (7)(8)(9)(10)(11)(12)(13)(14)(15)(16), hematopoietic stem cells (HSCs) from two organs (4,17), MSCs (4,18), EpSC (4), limbal epithelial stem cells (LSCs) (19), endothelial progenitor cells (EPCs) from different organs (3,4,(20)(21)(22)(23)(24)(25), supra-adventitial adipose stromal cells (SA-ASCs) (3,(22)(23)(24)(25), adipose pericytes (pericyte) (3,(22)(23)(24)(25), and finally cancer stem cells (CSCs) (26).…”

“…VI (4) VII (19) VIII (20,21) IX (3,(22)(23)(24)(25) X (4) XI (3,(22)(23)(24)(25) XII (3,(22)(23)(24)(25) XIII ( (17) IV (4) V (4,18) VI (4) VII (19) VIII (20,21) IX (3,(22)(23)(24)(25) X (4) XI (3,(22)(23)(24)(25) XII (3,(22)(23)(24)(25) XIII (26) RHAMM/HMMR marker expression in the same stem cell lineage obtained from various tissues may indicate differences in their pluripotency. It is also possible that stem cell marker expression pattern and levels depend on respective cell cycle phases.…”