Abstract
The human umbilical cord (UC) is an attractive source of mesenchymal stem cells (MSCs) with unique advantages over other MSC sources. They have been isolated from different compartments of the UC but there has been no rigorous comparison to identify the compartment with the best clinical utility. We compared the histology, fresh and cultured cell numbers, morphology, proliferation, viability, stemness characteristics and differentiation potential of cells from the amnion (AM), subamnion (SA), perivascular (PV), Wharton’s jelly (WJ) and mixed cord (MC) of five UCs. The WJ occupied the largest area in the UC from which 4.61 ± 0.57 x 106 /cm fresh cells could be isolated without culture compared to AM, SA, PV and MC that required culture. The WJ and PV had significantly lesser CD40+ non-stem cell contaminants (26-27%) compared to SA, AM and MC (51-70%). Cells from all compartments were proliferative, expressed the typical MSC-CD, HLA, and ESC markers, telomerase, had normal karyotypes and differentiated into adipocyte, chondrocyte and osteocyte lineages. The cells from WJ showed significantly greater CD24+ and CD108+ numbers and fluorescence intensities that discriminate between MSCs and non-stem cell mesenchymal cells, were negative for the fibroblast-specific and activating-proteins (FSP, FAP) and showed greater osteogenic and chondrogenic differentiation potential compared to AM, SA, PV and MC. Cells from the WJ offer the best clinical utility as (i) they have less non-stem cell contaminants (ii) can be generated in large numbers with minimal culture avoiding changes in phenotype, (iii) their derivation is quick and easy to standardize, (iv) they are rich in stemness characteristics and (v) have high differentiation potential. Our results show that when isolating MSCs from the UC, the WJ should be the preferred compartment, and a standardized method of derivation must be used so as to make meaningful comparisons of data between research groups.
Highlights
Mesenchymal stem cells have been derived from various sources
Starting from the outside, the compartments could be identified as the amnion (AM), Fig 1. (A) (a-d) In situ H & E histological cross-sections of the human umbilical cord showing the various regions [Wharton’s jelly (WJ), perivascular area (PV), subamnion (SA), amnion (AM)] from which mesenchymal stem cells (MSCs) were derived
It has been reported that the derivation of MSCs from the SA requires several hours as the umbilical cord pieces need to be dissected into small pieces and incubated for 10 to 14 days to be established in culture [7,27,40]
Summary
Mesenchymal stem cells have been derived from various sources Those of fetal origin face ethical issues as they are isolated from human abortuses while MSCs from adult bone marrow and organs have the disadvantages of painful invasive harvest, limited cell numbers, diminishing stemness properties with age and short-lived stemness properties in vitro [1,2]. These disadvantages have prompted interest in the exploration of other sources. Several groups have categorized the human UC into various compartments such as (i) the amniotic epithelial membrane (AM) (ii) subamnion or ‘cord lining’ (SA) (iii) intervascular Wharton’s jelly (WJ) and (iv) perivascular region (PV) surrounding the umbilical blood vessels [5,10]
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