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An efficient protocol to generate placental chorionic plate-derived mesenchymal stem cells with superior proliferative and immunomodulatory properties.

ABSTRACT: BACKGROUND:Placenta-derived MSCs (P-MSCs) represent a promising tool for cell-based therapeutic applications. However, the increasing demand for P-MSCs in clinical trials makes high quality and large number of P-MSCs mandatory. Here, we aim to develop an efficient protocol for P-MSC isolation and culture. METHODS:The modified explant culture (MEC) method by combining an initial mild enzymatic reaction with the subsequent explant culture was developed to simultaneously produce various P-MSCs from the different regions of the placenta in serum-free medium (SFM). Its isolation efficiencies, cell yield, and proliferative capacity were compared with the conventional explant culture (EC) method. Furthermore, we determined whether functional properties of P-MSCs are affected by the used tissue-harvesting sites in terms of their proliferation, migration, and the immunomodulatory effect on macrophage. RESULTS:The MEC method achieved higher yield and shorter time in primary cell confluence in SFM compared with the conventional method. The harvested cells possessed the MSC characteristics and demonstrated significantly stronger proliferation ability. Importantly, MSCs derived from chorionic plate (CP-MSCs) were found to exhibit superior properties to the other P-MSCs in proliferation and migration capacity, maintaining the fetal origin over serial passages. Notably, CP-MSCs show stronger ability in regulating macrophage polarization from M1 to M2. CONCLUSION:Our study developed an efficient and high-yield technique to produce high-quality P-MSCs from the placenta, hence serving as an optimal source of MSCs for clinical application.

SUBMITTER: Huang Q

PROVIDER: S-EPMC6796371 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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An efficient protocol to generate placental chorionic plate-derived mesenchymal stem cells with superior proliferative and immunomodulatory properties.

Huang Qilin Q Yang Yi Y Luo Chen C Wen Yi Y Liu Ruohong R Li Shuai S Chen Tao T Sun Hongyu H Tang Lijun L

Stem cell research & therapy 20191017 1

<h4>Background</h4>Placenta-derived MSCs (P-MSCs) represent a promising tool for cell-based therapeutic applications. However, the increasing demand for P-MSCs in clinical trials makes high quality and large number of P-MSCs mandatory. Here, we aim to develop an efficient protocol for P-MSC isolation and culture.<h4>Methods</h4>The modified explant culture (MEC) method by combining an initial mild enzymatic reaction with the subsequent explant culture was developed to simultaneously produce vari ...[more]

PMID: 31623677

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Project description:BackgroundNo regenerative approach has thus far been shown to be effective in skeletal muscle injuries, despite their high frequency and associated functional deficits. We sought to address surgical trauma-related muscle injuries using local intraoperative application of allogeneic placenta-derived, mesenchymal-like adherent cells (PLX-PAD), using hip arthroplasty as a standardized injury model, because of the high regenerative and immunomodulatory potency of this cell type.MethodsOur pilot phase I/IIa study was prospective, randomized, double blind, and placebo-controlled. Twenty patients undergoing hip arthroplasty via a direct lateral approach received an injection of 3.0 × 108 (300 M, n = 6) or 1.5 × 108 (150 M, n = 7) PLX-PAD or a placebo (n = 7) into the injured gluteus medius muscles.ResultsWe did not observe any relevant PLX-PAD-related adverse events at the 2-year follow-up. Improved gluteus medius strength was noted as early as Week 6 in the treatment-groups. Surprisingly, until Week 26, the low-dose group outperformed the high-dose group and reached significantly improved strength compared with placebo [150 M vs. placebo: P = 0.007 (baseline adjusted; 95% confidence interval 7.6, 43.9); preoperative baseline values mean ± SE: placebo: 24.4 ± 6.7 Nm, 150 M: 27.3 ± 5.6 Nm], mirrored by an increase in muscle volume [150 M vs. placebo: P = 0.004 (baseline adjusted; 95% confidence interval 6.0, 30.0); preoperative baseline values GM volume: placebo: 211.9 ± 15.3 cm3 , 150 M: 237.4 ± 27.2 cm3 ]. Histology indicated accelerated healing after cell therapy. Biomarker studies revealed that low-dose treatment reduced the surgery-related immunological stress reaction more than high-dose treatment (exemplarily: CD16+ NK cells: Day 1 P = 0.06 vs. placebo, P = 0.07 vs. 150 M; CD4+ T-cells: Day 1 P = 0.04 vs. placebo, P = 0.08 vs. 150 M). Signs of late-onset immune reactivity after high-dose treatment corresponded to reduced functional improvement.ConclusionsAllogeneic PLX-PAD therapy improved strength and volume of injured skeletal muscle with a reasonable safety profile. Outcomes could be positively correlated with the modulation of early postoperative stress-related immunological reactions.

Project description:BackgroundMesenchymal stem cells (MSCs) have emerged as a promising regenerative tool, owing mainly to their multi-differentiation potential and immunosuppressive capacity. When compared with MSCs classically derived from the adult bone marrow (BM), MSCs of neonatal origins exhibit superior proliferation ability, lower immunogenicity, and possible lower incorporated mutation; hence, they are considered as an alternative source for clinical use. Several researches have focused on the biological differences among some neonatal MSCs cultured in serum-containing medium (SCM). However, since it has been reported that MSCs possess different biological characteristics when cultured in serum-free medium (SFM), these comparative studies in SCM cannot exactly represent the results under the serum-free Good Manufacturing Practice (GMP) standard.MethodsHere, MSCs were isolated from three neonatal tissues, namely amniotic membrane (AM), umbilical cord (UC), and chorionic plate (CP), from the same donor, and their morphologies, immunophenotypes, trilineage differentiation potentials, global gene expression patterns, and proliferation abilities were systematically compared under chemical-defined SFM.ResultsOur study demonstrated that these three neonatal MSCs exhibited a similar morphology and immunophenotypic pattern but various mesodermal differentiation potentials under SFM: amniotic membrane-derived MSCs showed a higher rate for osteogenic differentiation; chorionic plate-derived MSCs presented better adipogenic induction efficiency; and all these three neonatal MSCs exhibited similar chondrogenic potential. Moreover, by the analysis of global gene expression patterns, we speculated a possible higher proliferation ability of CP-MSCs in SFM, and we subsequently validated this conjecture.ConclusionsCollectively, these results suggest that MSCs of different neonatal origins possess different biological features in SFM and thus may represent an optimal choice for different clinical applications.

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An efficient protocol to generate placental chorionic plate-derived mesenchymal stem cells with superior proliferative and immunomodulatory properties.

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An efficient protocol to generate placental chorionic plate-derived mesenchymal stem cells with superior proliferative and immunomodulatory properties.

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