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Comparative Characterization of Cells from the Various Compartments of the Human Umbilical Cord Shows that the Wharton's Jelly Compartment Provides the Best Source of Clinically Utilizable Mesenchymal Stem Cells.

Subramanian A, Fong CY, Biswas A, Bongso A - PLoS ONE (2015)

Bottom Line: The WJ and PV had significantly lesser CD40+ non-stem cell contaminants (26-27%) compared to SA, AM and MC (51-70%).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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynaecology, Yong Loo Lin School of Medicine, National University Health System, National University of Singapore, Kent Ridge, Singapore, 119228, Singapore.

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.

No MeSH data available.


Related in: MedlinePlus

(A-F) qRT-PCR for embryonic stem cell (ESC) pluripotent genomic markers in cells derived from the WJ, PV, SA, AM and MC.Cells derived from WJ expressed significantly greater levels of the pluripotency genes OCT1, OCT4A, OCT4B, NANOG and SOX2 compared to cells from SA, AM and MC. (*p<0.05). (G-H) Cells from SA, AM and MC showed significantly greater levels of expression of the fibroblast-related genes FAP and FSP compared to WJ and PV (*p<0.05).
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pone.0127992.g006: (A-F) qRT-PCR for embryonic stem cell (ESC) pluripotent genomic markers in cells derived from the WJ, PV, SA, AM and MC.Cells derived from WJ expressed significantly greater levels of the pluripotency genes OCT1, OCT4A, OCT4B, NANOG and SOX2 compared to cells from SA, AM and MC. (*p<0.05). (G-H) Cells from SA, AM and MC showed significantly greater levels of expression of the fibroblast-related genes FAP and FSP compared to WJ and PV (*p<0.05).

Mentions: Immunohistochemical staining of cells from the various compartments of the UC were all positive for the pluripotency markers SSEA-3, SSEA-4, Tra-1-60 and Tra-1-81 (Fig 5A–5D). qRT-PCR analysis showed that the cells from all the compartments also expressed the pluripotency genes OCT1, OCT2, OCT4A, OCT4B, NANOG and SOX2 (Fig 6A–6F). However, the expression levels for OCT1, OCT4A, OCT4B, NANOG and SOX2 pluripotent genes were significantly lower for cells from the SA and AM compared to WJ (Fig 6A and 6C–6F). Also, OCT4A and OCT4B genes were significantly lower for cells from the MC compared to WJ (Fig 6C and 6D). The cells from the SA, AM and MC showed significantly high expression levels of the fibroblast-related genes such as fibroblast activating protein (FAP) and fibroblast stimulating protein (FSP) (0.86 fold to 100.64 fold) compared to cells from the WJ and PV (Fig 6G and 6H).


Comparative Characterization of Cells from the Various Compartments of the Human Umbilical Cord Shows that the Wharton's Jelly Compartment Provides the Best Source of Clinically Utilizable Mesenchymal Stem Cells.

Subramanian A, Fong CY, Biswas A, Bongso A - PLoS ONE (2015)

(A-F) qRT-PCR for embryonic stem cell (ESC) pluripotent genomic markers in cells derived from the WJ, PV, SA, AM and MC.Cells derived from WJ expressed significantly greater levels of the pluripotency genes OCT1, OCT4A, OCT4B, NANOG and SOX2 compared to cells from SA, AM and MC. (*p<0.05). (G-H) Cells from SA, AM and MC showed significantly greater levels of expression of the fibroblast-related genes FAP and FSP compared to WJ and PV (*p<0.05).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4464659&req=5

pone.0127992.g006: (A-F) qRT-PCR for embryonic stem cell (ESC) pluripotent genomic markers in cells derived from the WJ, PV, SA, AM and MC.Cells derived from WJ expressed significantly greater levels of the pluripotency genes OCT1, OCT4A, OCT4B, NANOG and SOX2 compared to cells from SA, AM and MC. (*p<0.05). (G-H) Cells from SA, AM and MC showed significantly greater levels of expression of the fibroblast-related genes FAP and FSP compared to WJ and PV (*p<0.05).
Mentions: Immunohistochemical staining of cells from the various compartments of the UC were all positive for the pluripotency markers SSEA-3, SSEA-4, Tra-1-60 and Tra-1-81 (Fig 5A–5D). qRT-PCR analysis showed that the cells from all the compartments also expressed the pluripotency genes OCT1, OCT2, OCT4A, OCT4B, NANOG and SOX2 (Fig 6A–6F). However, the expression levels for OCT1, OCT4A, OCT4B, NANOG and SOX2 pluripotent genes were significantly lower for cells from the SA and AM compared to WJ (Fig 6A and 6C–6F). Also, OCT4A and OCT4B genes were significantly lower for cells from the MC compared to WJ (Fig 6C and 6D). The cells from the SA, AM and MC showed significantly high expression levels of the fibroblast-related genes such as fibroblast activating protein (FAP) and fibroblast stimulating protein (FSP) (0.86 fold to 100.64 fold) compared to cells from the WJ and PV (Fig 6G and 6H).

Bottom Line: The WJ and PV had significantly lesser CD40+ non-stem cell contaminants (26-27%) compared to SA, AM and MC (51-70%).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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynaecology, Yong Loo Lin School of Medicine, National University Health System, National University of Singapore, Kent Ridge, Singapore, 119228, Singapore.

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.

No MeSH data available.


Related in: MedlinePlus