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Nanoparticle labeling identifies slow cycling human endometrial stromal cells.

Xiang L, Chan RW, Ng EH, Yeung WS - Stem Cell Res Ther (2014)

Bottom Line: It remains unclear whether slow-cycling cells exist in the human endometrium.They also differentiate into multiple mesenchymal lineages and the expression of lineage specific markers was lower than that of non-FPC.In summary, nanoparticle labeling is a promising tool for the identification of putative somatic stem or progenitor cells when their surface markers are undefined.

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: Evidence suggests that the human endometrium contains stem or progenitor cells that are responsible for its remarkable regenerative capability. A common property of somatic stem cells is their quiescent state. It remains unclear whether slow-cycling cells exist in the human endometrium. We hypothesized that the human endometrium contains a subset of slow-cycling cells with somatic stem cell properties. Here, we established an in vitro stem cell assay to isolate human endometrial-derived mesenchymal stem-like cells (eMSC).

Methods: Single-cell stromal cultures were initially labeled with fluorescent nanoparticles and a small population of fluorescent persistent cells (FPC) remained after culture of 21 days. Two populations of stromal cells, namely FPC and non-FPC were sorted.

Results: Quantitative analysis of functional assays demonstrated that the FPC had higher colony forming ability, underwent more rounds of self-renewal and had greater enrichment of phenotypically defined prospective eMSC markers: CD146+/CD140b+ and W5C5+ than the non-FPC. They also differentiate into multiple mesenchymal lineages and the expression of lineage specific markers was lower than that of non-FPC. The FPC exhibit low proliferation activities. A proliferation dynamics study revealed that more FPC had a prolonged G1 phase.

Conclusions: With this study we present an efficient method to label and isolate slow-proliferating cells obtained from human endometrial stromal cultures without genetic modifications. The FPC population could be easily maintained in vitro and are of interest for tissue-repair and engineering perspectives. In summary, nanoparticle labeling is a promising tool for the identification of putative somatic stem or progenitor cells when their surface markers are undefined.

No MeSH data available.


Related in: MedlinePlus

Serial passage, clonogenicity and phenotyping of endometrial FPC and non-FPC post labeled day 21. (A) Rate of serial passage is shown for both large and small CFU of FPC (white) and non-FPC (black). (B) Percentage of large and small CFU at each passage of serial cloning for FPC (white) and non-FPC (black). Percentage of (C) CD146+/CD140b+ and (D) W5C5+ cells from FPC and non-FPC populations. Bar represents the mean. Representative dot-plots for co-staining of CD146/CD140b and single staining of W5C5. Single parameter histograms for individual markers CD146-FITC, CD140b-PE and W5C5-PE. Grey line indicates background fluorescence with isotype matched IgG control Results are reported as mean ± SEM, n = 7, *P < 0.05, **P < 0.01, ***P < 0.001. CFU, colony-forming units; FPC, fluorescent persistent cells; SEM, standard error of the mean.
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Figure 3: Serial passage, clonogenicity and phenotyping of endometrial FPC and non-FPC post labeled day 21. (A) Rate of serial passage is shown for both large and small CFU of FPC (white) and non-FPC (black). (B) Percentage of large and small CFU at each passage of serial cloning for FPC (white) and non-FPC (black). Percentage of (C) CD146+/CD140b+ and (D) W5C5+ cells from FPC and non-FPC populations. Bar represents the mean. Representative dot-plots for co-staining of CD146/CD140b and single staining of W5C5. Single parameter histograms for individual markers CD146-FITC, CD140b-PE and W5C5-PE. Grey line indicates background fluorescence with isotype matched IgG control Results are reported as mean ± SEM, n = 7, *P < 0.05, **P < 0.01, ***P < 0.001. CFU, colony-forming units; FPC, fluorescent persistent cells; SEM, standard error of the mean.

Mentions: The self-renewal ability of endometrial stromal FPC large CFU at chase-D21 (4.2 ± 0.4) was significantly higher than that of the non-FPC large CFU (2.1 ± 0.1, P < 0.01, n = 7; Figure 3A) when more samples were analyzed. The self-renewal activity was also higher for the endometrial stromal FPC small CFU (1.50 ± 0.13) than for the non-FPC small CFU (1.1 ± 0.1, P < 0.05).


Nanoparticle labeling identifies slow cycling human endometrial stromal cells.

Xiang L, Chan RW, Ng EH, Yeung WS - Stem Cell Res Ther (2014)

Serial passage, clonogenicity and phenotyping of endometrial FPC and non-FPC post labeled day 21. (A) Rate of serial passage is shown for both large and small CFU of FPC (white) and non-FPC (black). (B) Percentage of large and small CFU at each passage of serial cloning for FPC (white) and non-FPC (black). Percentage of (C) CD146+/CD140b+ and (D) W5C5+ cells from FPC and non-FPC populations. Bar represents the mean. Representative dot-plots for co-staining of CD146/CD140b and single staining of W5C5. Single parameter histograms for individual markers CD146-FITC, CD140b-PE and W5C5-PE. Grey line indicates background fluorescence with isotype matched IgG control Results are reported as mean ± SEM, n = 7, *P < 0.05, **P < 0.01, ***P < 0.001. CFU, colony-forming units; FPC, fluorescent persistent cells; SEM, standard error of the mean.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4230801&req=5

Figure 3: Serial passage, clonogenicity and phenotyping of endometrial FPC and non-FPC post labeled day 21. (A) Rate of serial passage is shown for both large and small CFU of FPC (white) and non-FPC (black). (B) Percentage of large and small CFU at each passage of serial cloning for FPC (white) and non-FPC (black). Percentage of (C) CD146+/CD140b+ and (D) W5C5+ cells from FPC and non-FPC populations. Bar represents the mean. Representative dot-plots for co-staining of CD146/CD140b and single staining of W5C5. Single parameter histograms for individual markers CD146-FITC, CD140b-PE and W5C5-PE. Grey line indicates background fluorescence with isotype matched IgG control Results are reported as mean ± SEM, n = 7, *P < 0.05, **P < 0.01, ***P < 0.001. CFU, colony-forming units; FPC, fluorescent persistent cells; SEM, standard error of the mean.
Mentions: The self-renewal ability of endometrial stromal FPC large CFU at chase-D21 (4.2 ± 0.4) was significantly higher than that of the non-FPC large CFU (2.1 ± 0.1, P < 0.01, n = 7; Figure 3A) when more samples were analyzed. The self-renewal activity was also higher for the endometrial stromal FPC small CFU (1.50 ± 0.13) than for the non-FPC small CFU (1.1 ± 0.1, P < 0.05).

Bottom Line: It remains unclear whether slow-cycling cells exist in the human endometrium.They also differentiate into multiple mesenchymal lineages and the expression of lineage specific markers was lower than that of non-FPC.In summary, nanoparticle labeling is a promising tool for the identification of putative somatic stem or progenitor cells when their surface markers are undefined.

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: Evidence suggests that the human endometrium contains stem or progenitor cells that are responsible for its remarkable regenerative capability. A common property of somatic stem cells is their quiescent state. It remains unclear whether slow-cycling cells exist in the human endometrium. We hypothesized that the human endometrium contains a subset of slow-cycling cells with somatic stem cell properties. Here, we established an in vitro stem cell assay to isolate human endometrial-derived mesenchymal stem-like cells (eMSC).

Methods: Single-cell stromal cultures were initially labeled with fluorescent nanoparticles and a small population of fluorescent persistent cells (FPC) remained after culture of 21 days. Two populations of stromal cells, namely FPC and non-FPC were sorted.

Results: Quantitative analysis of functional assays demonstrated that the FPC had higher colony forming ability, underwent more rounds of self-renewal and had greater enrichment of phenotypically defined prospective eMSC markers: CD146+/CD140b+ and W5C5+ than the non-FPC. They also differentiate into multiple mesenchymal lineages and the expression of lineage specific markers was lower than that of non-FPC. The FPC exhibit low proliferation activities. A proliferation dynamics study revealed that more FPC had a prolonged G1 phase.

Conclusions: With this study we present an efficient method to label and isolate slow-proliferating cells obtained from human endometrial stromal cultures without genetic modifications. The FPC population could be easily maintained in vitro and are of interest for tissue-repair and engineering perspectives. In summary, nanoparticle labeling is a promising tool for the identification of putative somatic stem or progenitor cells when their surface markers are undefined.

No MeSH data available.


Related in: MedlinePlus