Phenotypic and cytogenetic characterization of expanded adipose derived stem cells

Introduction: Human adipose derived stem cells (hASCs) have great potential for regenerative medicine. The demand for hASCs, especially in the development of off-the-shelf products, is increasing. Although the initial receipt of hASCs was relatively limited, there is now greater interest and also awareness that in vitro expansion of hASCs be further explored. The purpose of this study was to assess the integrity of mesenchymal cell characteristics and the mutant capability of chromosome number on hASCs undergoing in vitro expansion. Methods: In this study, three hASC samples from three Vietnamese people were collected and proliferated in MSCCult medium (Regemedlab, Ho Chi Minh City, Viet Nam) to the 5th cell passage. Next, hASCs were evaluated for change of mesenchymal stem cells (MSCs) characteristics including shape, immunophenotype (CD14, CD34, CD44, CD73, CD90, and/or CD166), and trilineage differentiation ability. Finally, the number of chromosomes after passages 1, 3, and 5 was evaluated by karyotyping technique. Results: The results showed that after five passages of culture, hASCs preserved the characteristic shape of MSCs, high expression of mesenchymal markers (e.g. CD44, CD73, CD90, and CD166). However, the cells also maintained their differentiation capacity to develop into various tissues such as bone, cartilage, and fat. The hASCs showed no mutation in the number of chromosomes. However, markers of hematopoietic cells (such as CD14 and CD34) exhibited heterogeneous changes between the samples during proliferation. Conclusion: In conclusion, at passage 5, hASCs retained the integrity of MSC features and there was no mutation discovered in the number of chromosomes. However, further evaluation is needed to conclude that the use of cultured cells in treatment is effective and safe.