Stem Cell Identification and Clinical Practice

Abstract

Stem cells are non-differentiated cells that are capable of proliferation at regular intervals, differentiation into cells in need, self-regenerating
and repairing the degenerative tissues in an organism. In this study, we aimed to isolate mesenchymal stem cells from adipose tissue (ADSCs)
and induce the differentiation of ADSCs into osteoblast, chondroblast and adipocyte cell types, and to characterize these cells using positive
and negative stem cell markers in order to prepare ADSCs for clinical application.
For this purpose, no chemical-based enzymes were used in ADSCs production. ADSCs were isolated from live tissue using a primary cell
culture technique and cells were propogated for up to three weeks through third passage. Third passage cells were differentiated into osteoblast,
chondroblast and adipocytes. For histological identification of multi-lineage differentiation of cells, staining techniques including Alizarin Red,
Alcian Blue and Oil Red O were applied. Before transplantation, ADSCs were immunohistochemically characterized by positive expression
of CD 90, CD 105 and negative expression of CD 45, CD11b. For the transplantation of stem cells 3rd passage cells were subcultured by
trypsinization. Trypan blue staining method was used for the analysis of living cells. Cells were counted by invert microscope under a 20×
objective. For the transplantation of stem cells, the contents of the autologous blood serum and stem cells were transferred into the region of
the operation at a dose of 1 × 106 cells / 100 μl by the insulin injector.
In conclusion, we aim to contribute to clinical stem cell studies and literature knowledge by isolating MSCs from adipose tissue followed by
differentiation and characterization of these cells using various histological and immunohistochemical techniques, as well as by determining
suitable stem cell dose for clinical applications.