The Relative Functionality of Freshly Isolated and Cryopreserved Human Adipose-Derived Stromal/Stem Cells

Abstract

The capability of multipotent mesenchymal stem cells to maintain cell viability, phenotype and differentiation ability upon thawing is critical if they are to be banked and used for future therapeutic purposes. In the present study, we examined the effect of 9-10 months of cryostorage on the morphology, immunophenotype, colony-forming unit (CFU) and differentiation capacity of fresh and cryopreserved human adipose-derived stromal/stem cells (ASCs) from the same donors. Cryopreservation did not reduce the CFU frequency and the expression levels of CD29, CD73, CD90 and CD105 remained unchanged with the exception of CD34 and CD45; however, the differentiation capacity of cryopreserved ASCs relative to fresh cells was significantly reduced. While our findings suggest that future studies are warranted to improve cryopreservation methods and agents, cryopreserved ASCs retain sufficient features to ensure their practical utility for both research and clinical applications.

hASC and DFAT, Multipotent Stem Cells for Regenerative Medicine: A Comparison of Their Potential Differentiation In Vitro

“…This tissue contains cells that express specific markers of mesenchymal stem cells, termed hASC. Literature data about the expression of CD34 in hASC are conflicting: it is positive according to some studies [ 29 , 30 ] and negative according to others [ 31 , 32 ]. In particular, it has been suggested that some hematopoietic cells (CD34+) are present in earlier passages and lost in the next steps.…”

“…In particular, it has been suggested that some hematopoietic cells (CD34+) are present in earlier passages and lost in the next steps. hASCs are positive for mesenchymal stem cell markers as CD13, CD73, CD90, and CD105, and negative for CD14, CD34 [ 31 , 32 ], and CD45, according to Tocci et al (2003) [ 33 ]. The cells obtained from mature adipocytes and maintained in “ceiling mode” start a gradual process of dedifferentiation, highlighted by positive markers of mesenchymal stem cells [ 28 , 34 ].…”

Human Adipose Derived Cells in Two- and Three-Dimensional Cultures: Functional Validation of an In Vitro Fat Construct

“…While a number of studies have demonstrated the feasibility of cryopreserving intact adipose tissue fragments for extended periods of time, only~50% of the SVF cell population within these tissues remain viable [23]. This contrasts with cryopreservation of isolated SVF cells and ASC which retain postthaw viability levels of 80% or better, depending on the isolation process and cryopreservation agent [23][24][25][26][27]. It is noteworthy that in the current study, the SVF cell immunophenotype differed between freshly prepared and cryopreserved/thawed cells.…”

Multipotency and secretome: the mechanisms behind the regenerative potential of adipose-derived stem cells

“…According to Shah et al [21] colony forming potential and cell marker expression of ASCs except CD34 and CD45 did not change with cryopreservation; however, their differentiation potential was significantly reduced. Further investigation on this subject is necessary.…”