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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

Clonogenicity and self- renewal ability of stromal FPC and non-FPC post-labeled day 15 and 21. (A – D) Cloning efficiency of endometrial stromal cells post-labeled with nanoparticles at day 15 (white bars) and 21 (grey bars). Cloning efficiency of large CFU (A) FPC and (B) non-FPC. Cloning efficiency of small CFU (C) FPC and (D) non-FPC. (E – H) Self-renewal activity of endometrial stromal cells post-labeled with nanoparticles at day 15 (white bars) and 21 (grey bars) using serial cloning assay. Large CFU self-renewal ability of (E) FPC and (F) non-FPC. Small CFU self-renewal ability of (G) FPC and (H) non-FPC Results reported as means ± SEM, n = 4, *, a, bP <0.05. CFU, colony-forming units; FPC, fluorescent persistent cells; SEM, standard error of the mean.
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Figure 2: Clonogenicity and self- renewal ability of stromal FPC and non-FPC post-labeled day 15 and 21. (A – D) Cloning efficiency of endometrial stromal cells post-labeled with nanoparticles at day 15 (white bars) and 21 (grey bars). Cloning efficiency of large CFU (A) FPC and (B) non-FPC. Cloning efficiency of small CFU (C) FPC and (D) non-FPC. (E – H) Self-renewal activity of endometrial stromal cells post-labeled with nanoparticles at day 15 (white bars) and 21 (grey bars) using serial cloning assay. Large CFU self-renewal ability of (E) FPC and (F) non-FPC. Small CFU self-renewal ability of (G) FPC and (H) non-FPC Results reported as means ± SEM, n = 4, *, a, bP <0.05. CFU, colony-forming units; FPC, fluorescent persistent cells; SEM, standard error of the mean.

Mentions: The cloning efficiency was determined for the large and small CFU from both populations. The percentage of FPC that formed large CFU was 0.3 ± 0.1% (n = 4) at chase-D15 and 0.6 ± 0.2% (n = 4) at chase-D21 (Figure 2A). Although there was a trend of higher large CFU forming ability with increased duration of chase, the increase did not reach statistical significance (P = 0.34). The corresponding cloning efficiencies of the non-FPC large CFU were 0.3 ± 0.1% and 0.4 ± 0.1%, respectively (Figure 2B). The cloning efficiencies for FPC small CFU at chase-D15 (1.9 ± 0.3%) and chase-D21 (1.7 ± 0.5%) were similar (Figure 2C). The cloning efficiencies of non-FPC small CFU were also similar (D15: 2.7 ± 0.2%; D21: 3.2 ± 1.2%; Figure 2D). There was no statistical difference in the formation of small CFU for FPC (P = 0.69) and non-FPC (P = 1.00) at the two chase periods.


Nanoparticle labeling identifies slow cycling human endometrial stromal cells.

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

Clonogenicity and self- renewal ability of stromal FPC and non-FPC post-labeled day 15 and 21. (A – D) Cloning efficiency of endometrial stromal cells post-labeled with nanoparticles at day 15 (white bars) and 21 (grey bars). Cloning efficiency of large CFU (A) FPC and (B) non-FPC. Cloning efficiency of small CFU (C) FPC and (D) non-FPC. (E – H) Self-renewal activity of endometrial stromal cells post-labeled with nanoparticles at day 15 (white bars) and 21 (grey bars) using serial cloning assay. Large CFU self-renewal ability of (E) FPC and (F) non-FPC. Small CFU self-renewal ability of (G) FPC and (H) non-FPC Results reported as means ± SEM, n = 4, *, a, bP <0.05. 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 2: Clonogenicity and self- renewal ability of stromal FPC and non-FPC post-labeled day 15 and 21. (A – D) Cloning efficiency of endometrial stromal cells post-labeled with nanoparticles at day 15 (white bars) and 21 (grey bars). Cloning efficiency of large CFU (A) FPC and (B) non-FPC. Cloning efficiency of small CFU (C) FPC and (D) non-FPC. (E – H) Self-renewal activity of endometrial stromal cells post-labeled with nanoparticles at day 15 (white bars) and 21 (grey bars) using serial cloning assay. Large CFU self-renewal ability of (E) FPC and (F) non-FPC. Small CFU self-renewal ability of (G) FPC and (H) non-FPC Results reported as means ± SEM, n = 4, *, a, bP <0.05. CFU, colony-forming units; FPC, fluorescent persistent cells; SEM, standard error of the mean.
Mentions: The cloning efficiency was determined for the large and small CFU from both populations. The percentage of FPC that formed large CFU was 0.3 ± 0.1% (n = 4) at chase-D15 and 0.6 ± 0.2% (n = 4) at chase-D21 (Figure 2A). Although there was a trend of higher large CFU forming ability with increased duration of chase, the increase did not reach statistical significance (P = 0.34). The corresponding cloning efficiencies of the non-FPC large CFU were 0.3 ± 0.1% and 0.4 ± 0.1%, respectively (Figure 2B). The cloning efficiencies for FPC small CFU at chase-D15 (1.9 ± 0.3%) and chase-D21 (1.7 ± 0.5%) were similar (Figure 2C). The cloning efficiencies of non-FPC small CFU were also similar (D15: 2.7 ± 0.2%; D21: 3.2 ± 1.2%; Figure 2D). There was no statistical difference in the formation of small CFU for FPC (P = 0.69) and non-FPC (P = 1.00) at the two chase periods.

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