Abstract
IntroductionUmbilical cord blood (UCB)-derived mesenchymal stem cells (MSCs) are self-renewing multipotent progenitors with the potential to differentiate into multiple lineages of mesoderm, in addition to generating ectodermal and endodermal lineages by crossing the germline barrier. In the present study we have investigated the ability of UCB-MSCs to generate neurons, since we were able to observe varying degrees of neuronal differentiation from a few batches of UCB-MSCs with very simple neuronal induction protocols whereas other batches required extensive exposure to combination of growth factors in a stepwise protocol. Our hypothesis was therefore that the human UCB-MSCs would contain multiple types of progenitors with varying neurogenic potential and that the ratio of the progenitors with high and low neurogenic potentials varies in different batches of UCB.MethodsIn total we collected 45 UCB samples, nine of which generated MSCs that were further expanded and characterized using immunofluorescence, fluorescence-activated cell sorting and RT-PCR analysis. The neuronal differentiation potential of the UCB-MSCs was analyzed with exposure to combination of growth factors.ResultsWe could identify two different populations of progenitors within the UCB-MSCs. One population represented progenitors with innate neurogenic potential that initially express pluripotent stem cell markers such as Oct4, Nanog, Sox2, ABCG2 and neuro-ectodermal marker nestin and are capable of expanding and differentiating into neurons with exposure to simple neuronal induction conditions. The remaining population of cells, typically expressing MSC markers, requires extensive exposure to a combination of growth factors to transdifferentiate into neurons. Interesting to note was that both of these cell populations were positive for CD29 and CD105, indicating their MSC lineage, but showed prominent difference in their neurogenic potential.ConclusionOur results suggest that the expanded UCB-derived MSCs harbor a small unique population of cells that express pluripotent stem cell markers along with MSC markers and possess an inherent neurogenic potential. These pluripotent progenitors later generate cells expressing neural progenitor markers and are responsible for the instantaneous neuronal differentiation; the ratio of these pluripotent marker expressing cells in a batch determines the innate neurogenic potential.
Highlights
Umbilical cord blood (UCB)-derived mesenchymal stem cells (MSCs) are self-renewing multipotent progenitors with the potential to differentiate into multiple lineages of mesoderm, in addition to generating ectodermal and endodermal lineages by crossing the germline barrier
Our initial interest was to isolate MSCs from UCB and check their potential to transdifferentiate into neurons, but surprisingly we found significant variations between the isolated MSCs from different batches - this led us to further characterize these human umbilical cord blood (hUCB)-MSCs
We observed a very significantly high percentage (30.08 ± 4.26, P < 0.001) of b-III tubulin-expressing cells in hUCBMSC SCB4 and moderately high neuronal differentiation in hUCB-MSC SCB10 (9.37 ± 1.84), compared with the RCB 2080 cell line (1.22 ± 0.61; Figure 7B to 7M, N). These results further indicate that the RCB 2080 cell line, which has a typical MSC phenotype, may be transdifferentiating alone as a result of pressure from the combination of growth factors, whereas the hUCB-MSC SCB4 and hUCB-MSC SCB10 cells co-expressing MSC and neural progenitor markers, with inherent neurogenic potential, appear to differentiate into neurons without much pressure or induction with the combination of growth factors
Summary
Umbilical cord blood (UCB)-derived mesenchymal stem cells (MSCs) are self-renewing multipotent progenitors with the potential to differentiate into multiple lineages of mesoderm, in addition to generating ectodermal and endodermal lineages by crossing the germline barrier. The collection of mesenchymal stem cells (MSCs) from UCB that is discarded at the time of birth is an easier, less expensive and non-invasive method than collecting MSCs from bone marrow aspirates [2]. These MSCs attract special interest due to these specific advantages over embryonic and adult stem cell counterparts, since there are no ethical issues associated with UCB. Another important characteristic of UCB-MSCs is that they are less immunogenic, and do not elicit the proliferative response of allogeneic lymphocytes in vitro [3]. Cells derived from the UCB elicit a lower incidence of graft rejection and post-transplant infections compared with other sources [4]
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