Homogeneity evaluation of mesenchymal stem cells based on electrotaxis analysis

Abstract

Stem cell therapy that can restore function to damaged tissue, avoid host rejection and reduce inflammation throughout body without use of immunosuppressive drugs. The established methods were used to identify and to isolate specific stem cell markers by FACS or by immunomagnetic cell separation. The procedures for distinguishing population of stem cells took a time and needed many preparations. Here we suggest an electrotaxis analysis as a new method to evaluate the homogeneity of mesenchymal stem cells which can observe the stem cell population in culture condition and wide use to various types of stem cells. Human mesenchymal stem cell, adipose derived stem cell, tonsil derived stem cell and osteogenic differentiated cells migrated toward anode but the migration speed of differentiated cells was significantly decreased versus that of stem cells. In mixture of stem cells and differentiated cells condition, we identified that the ratio of stem cell versus differentiated cell was matched with the homogeneity evaluation data of stem cells based on electrotaxis analysis. As a result, our evaluation tool has the possibility of the wide use to stem cell homogeneity evaluation and might be used as the stem cell quality control during stem cell culture without any additional antibodies.

Proliferative, Migratory, and Transition Properties Reveal Metastate of Human Amnion Cells

“…After 22 hours, dominance of vimentin-positive cells with mesenchymal morphologic features at the leading edge were seen ( Figure 2C). Mesenchymal cells are more migratory, 35 and the CK-18/vimentin staining pattern suggests that EMT promoted cellular migration to heal the wound. Wounds were healed within 38 hours, and examination of these sites showed dominance of intermediate filament CK-18 þ cells with a morphologic finding that indicated MET ( Figure 2C).…”

Pulsed Electrical Stimulation Enhances Consistency of Directional Migration of Adipose-Derived Stem Cells

“…In other words, although cell groups were exposed to ES for an equivalent time period (continuous ES for 6 h as opposed to two repetitions of 3 h ES each), the repetition of shorter bursts of ES was more effective in maintaining the cell migration direction without causing cell damage. We had previously shown that human tonsil-, adipose tissue-, and bone marrowderived mesenchymal stem cells (MSC) migrate directly toward the anode when exposed to an electric current [24,25]. Other published studies have also indicated that either direct or alternating current stimulate the directional migration of cells toward a specific cathodal or anodal orientation [23,[36][37][38][39][40].…”

“…We had previously shown that human tonsil-, adipose tissue-, and bone marrow-derived mesenchymal stem cells (MSC) migrate directly toward the anode when exposed to an electric current [ 24 , 25 ]. Other published studies have also indicated that either direct or alternating current stimulate the directional migration of cells toward a specific cathodal or anodal orientation [ 23 , 36 , 37 , 38 , 39 , 40 ].…”

“…3.8 software (Live Cell Instrument, Inc., Seoul, Korea), and equipped with an inverted microscope (Olympus Corporation, Tokyo, Japan). The electrotaxis chamber consisted of a chamber base on which a cell-seeded glass slide was placed, followed by a silicon gasket and finally the chamber top, connected by two 2% agar-salt bridges in Steinberg’s solution ( Figure 1 ) [ 24 ]. Human ADSCs (Lonza, Basel, Switzerland) were cultured in ADSC growth medium (ADSC-GM, Lonza) at 37 °C in a 5% CO 2 environment.…”

“…During electrotaxis, cells subjected to direct electric current will move preferentially toward either the anode or cathode, indicating that the effects of electric signals on cells are specific to different cell types and species [ 21 , 22 , 23 ]. Researchers have previously demonstrated that exogenous direct ES can effectively guide directional migration toward the anode of ADSCs, human mesenchymal stem cells (hMSCs), and tonsil derived MSCs [ 24 , 25 , 26 ]. In our previous studies, we showed that cells migrate toward the cathode or anode following Golgi apparatus (GA) polarization, in the same direction as the ES.…”