Dataset Information

Serum and xeno-free, chemically defined, no-plate-coating-based culture system for mesenchymal stromal cells from the umbilical cord.

ABSTRACT:

Objectives

Umbilical cord mesenchymal stromal cells (UCMSCs) can be considered to become a new gold standard for MSC-based therapies. A serum and xeno-free, chemically defined and no-plate-coating-based culture system will greatly facilitate development of robust, clinically acceptable bioprocesses for reproducibly generating quality-assured UCMSCs.

Materials and methods

In this study, we report for the first time, such a serum-free, xeno-free, completely chemically defined and no-plate-coating-based culture system for the isolation and expansion of UCMSCs, whose biological characteristics were evaluated and compared with serum-containing medium (SCM) methods.

Results

This culture system not only supported UCMSC primary cultures but also allowed for their expansion at low seeding density. Compared to SCM, UCMSCs in SFM exhibited (i) higher proliferative and colony-forming capacities; (ii) distinctly different morphologies; (iii) similar phenotype; (iv) similar pluripotency-associated marker expression; (v) superior osteogenic, but reduced adipogenic differentiation capacitities. In addition, UCMSCs cultured in SFM retained similar immunomodulatory properties to those in SCM.

Conclusions

Our findings demonstrate the feasibility of isolating and expanding UCMSCs in a completely serum-free, xeno-free, chemically defined and no-plate-coating-based culture system and represent an important step forward for development of robust, clinically acceptable bioprocesses for UCMSCs. Further, this provides a superior study platform for UCMSCs biology in a controlled environment.

SUBMITTER: Wu X

PROVIDER: S-EPMC6496339 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Serum and xeno-free, chemically defined, no-plate-coating-based culture system for mesenchymal stromal cells from the umbilical cord.

Wu Xiaoyun X Kang Huiyan H Liu Xuemin X Gao Jin J Zhao Kuijun K Ma Zhijie Z

Cell proliferation 20160804 5

<h4>Objectives</h4>Umbilical cord mesenchymal stromal cells (UCMSCs) can be considered to become a new gold standard for MSC-based therapies. A serum and xeno-free, chemically defined and no-plate-coating-based culture system will greatly facilitate development of robust, clinically acceptable bioprocesses for reproducibly generating quality-assured UCMSCs.<h4>Materials and methods</h4>In this study, we report for the first time, such a serum-free, xeno-free, completely chemically defined and no ...[more]

PMID: 27492579

Similar Datasets

Project description:Cell therapy is witnessing a notable shift toward cell-free treatments based on paracrine factors, in particular, towards small extracellular vesicles (sEV), that mimic the functional effect of the parental cells. While numerous sEV-based applications are currently in advanced preclinical stages, their promised translation depends on overcoming the manufacturing hurdles posed by the large-scale production of purified sEV. Unquestionably, the culture medium used with the parental cells plays a key role in the sEV's secretion rate and content. An essential requisite is the use of a serum-, xeno-, and blood-free medium to meet the regulatory entity requirements of clinical-grade sEV's production. Here, we evaluated OxiumTMEXO, a regulatory complying medium, with respect to production capacity and conservation of the EV's characteristics and functionality and the parental cell's phenotype and viability. A comparative study was established with standard DMEM and a commercially available culture medium developed specifically for sEV production. Under similar conditions, OxiumTMEXO displayed a three-fold increase of sEV secretion, with an enrichment of particles ranging between 51 and 200 nm. These results were obtained through direct quantification from the conditioned medium to avoid the isolation method's interference and variability and were compared to the two culture media under evaluation. The higher yield obtained was consistent with several harvest time points (2, 4, and 6 days) and different cell sources, incluiding umbilical cord-, menstrual blood-derived mesenchymal stromal cells and fibroblasts. Additionally, the stem cell phenotype and viability of the parental cell remained unchanged. Furthermore, OxiumTMEXO-sEV showed a similar expression pattern of the vesicular markers CD63, CD9, and CD81, with respect to sEV derived from the other conditions. The in vitro internalization assays in different target cell types and the pharmacokinetic profile of intraperitoneally administered sEV in vivo indicated that the higher EV production rate did not affect the uptake kinetics or the systemic biodistribution in healthy mice. In conclusion, the OxiumTMEXO medium sustains an efficient and robust production of large quantities of sEV, conserving the classic functional properties of internalization into acceptor target cells and biodistribution in vivo, supplying the amount and quality of EVs for the development of cell-free therapies.

Project description:Constructs currently used to repair or replace congenitally diseased pediatric heart valves lack a viable cell population capable of functional adaptation in situ, necessitating repeated surgical intervention. Heart valve tissue engineering (HVTE) can address these limitations by producing functional living tissue in vitro that holds the potential for somatic growth and remodelling upon implantation. However, clinical translation of HVTE strategies requires an appropriate source of autologous cells that can be non-invasively harvested from mesenchymal stem cell (MSC)-rich tissues and cultured under serum- and xeno-free conditions. To this end, we evaluated human umbilical cord perivascular cells (hUCPVCs) as a promising cell source for in vitro production of engineered heart valve tissue. The proliferative, clonogenic, multilineage differentiation, and extracellular matrix (ECM) synthesis capacities of hUCPVCs were evaluated in a commercial serum- and xeno-free culture medium (StemMACS™) on tissue culture polystyrene and benchmarked to adult bone marrow-derived MSCs (BMMSCs). Additionally, the ECM synthesis potential of hUCPVCs was evaluated when cultured on polycarbonate polyurethane anisotropic electrospun scaffolds, a representative biomaterial for in vitro HVTE. hUCPVCs had greater proliferative and clonogenic potential than BMMSCs in StemMACS™ (p < 0.05), without differentiation to osteogenic and adipogenic phenotypes associated with valve pathology. Furthermore, hUCPVCs cultured with StemMACS™ on tissue culture plastic for 14 days synthesized significantly more total collagen, elastin, and sulphated glycosaminoglycans (p < 0.05), the ECM constituents of the native valve, than BMMSCs. Finally, hUCPVCs retained their ECM synthesizing capacity after 14 and 21 days in culture on anisotropic electrospun scaffolds. Overall, our findings establish an in vitro culture platform that uses hUCPVCs as a readily-available and non-invasively sourced autologous cell population and a commercial serum- and xeno-free culture medium to increase the translational potential of future pediatric HVTE strategies. This study evaluated the proliferative, differentiation and extracellular matrix (ECM) synthesis capacities of human umbilical cord perivascular cells (hUCPVCs) when cultured in serum- and xeno-free media (SFM) against conventionally used bone marrow-derived MSCs (BMMSCs) and serum-containing media (SCM). Our findings support the use of hUCPVCs and SFM for in vitro heart valve tissue engineering (HVTE) of autologous pediatric valve tissue. Figure created with BioRender.com.

Dataset Information

Serum and xeno-free, chemically defined, no-plate-coating-based culture system for mesenchymal stromal cells from the umbilical cord.

Objectives

Materials and methods

Results

Conclusions

Publications

Serum and xeno-free, chemically defined, no-plate-coating-based culture system for mesenchymal stromal cells from the umbilical cord.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets