scholarly journals Efficacy of 3D Culture Priming is Maintained in Human Mesenchymal Stem Cells after Extensive Expansion of the Cells

Cells ◽  
2019 ◽  
Vol 8 (9) ◽  
pp. 1031 ◽  
Author(s):  
Thomas J. Bartosh ◽  
Joni H. Ylostalo
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
3D Culture ◽  
Human Mesenchymal Stem Cells ◽  
Early Passage ◽  
Animal Product ◽  
Large Numbers ◽  
Late Passage ◽  
Fetal Bovine ◽  
Free Media

The use of non-optimal preparations of mesenchymal stem cells (MSCs), such as extensively expanded cells, might be necessary to obtain the large numbers of cells needed for many clinical applications. We previously demonstrated that minimally expanded (early passage) MSCs can be pre-activated as spheroids to produce potentially therapeutic factors in 3D cultures. Here, we used extensively expanded (late passage) MSCs and studied their 3D-culture activation potential. MSCs were culture-expanded as 2D monolayers, and cells from various passages were activated by 3D culture in hanging drops with either fetal bovine serum (FBS)-containing media or a more clinically-applicable animal product-free (xeno-free) media. Gene expression analyses demonstrated that MSC spheroids prepared from passage 3, 5, and 7 cells were similar to each other but different from 2D MSCs. Furthermore, the expression of notable anti-inflammatory/immune-modulatory factors cyclooxygenase-2 (PTGS2), TNF alpha induced protein 6 (TNFAIP6), and stanniocalcin 1 (STC-1) were up-regulated in all spheroid preparations. This was confirmed by the detection of secreted prostaglandin E2 (PGE-2), tumor necrosis factor-stimulated gene 6 (TSG-6, and STC-1. This study demonstrated that extensively expanded MSCs can be activated in 3D culture through spheroid formation in both FBS-containing and xeno-free media. This work highlights the possibility of activating otherwise less useable MSC preparations through 3D culture generating large numbers of potentially therapeutic MSCs.

scholarly journals Ex VivoExpansion of Human Mesenchymal Stem Cells in Defined Serum-Free Media

2012 ◽  
Vol 2012 ◽  
pp. 1-21 ◽  
Author(s):  
Sunghoon Jung ◽  
Krishna M. Panchalingam ◽  
Lawrence Rosenberg ◽  
Leo A. Behie
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Therapeutic Potential ◽  
Culture Media ◽  
Human Mesenchymal Stem Cells ◽  
Efficient Production ◽  
Serum Free ◽  
Medium Development ◽  
Current Cell ◽  
Free Media

Human mesenchymal stem cells (hMSCs) are presently being evaluated for their therapeutic potential in clinical studies to treat various diseases, disorders, and injuries. To date, early-phase studies have indicated that the use of both autologous and allogeneic hMSCs appear to be safe; however, efficacy has not been demonstrated in recent late-stage clinical trials. Optimized cell bioprocessing protocols may enhance the efficacy as well as safety of hMSC therapeutics. Classical media used for generating hMSCs are typically supplemented with ill-defined supplements such as fetal bovine serum (FBS) or human-sourced alternatives. Ideally, culture media are desired to have well-defined serum-free formulations that support the efficient production of hMSCs while maintaining their therapeutic and differentiation capacity. Towards this objective, we review here current cell culture media for hMSCs and discuss medium development strategies.

Isolation of large numbers of human mesenchymal stem cells from discarded filters after bone marrow harvests: Implications for therapy

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 12501-12501
Author(s):  
A. S. Mageed ◽  
D. Pietryga ◽  
S. Gagin ◽  
R. West
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Mononuclear Cells ◽  
Human Mesenchymal Stem Cells ◽  
Published Data ◽  
Hla Dr ◽  
Large Numbers ◽  
Specific Antigens ◽  
Novel Strategy

12501 Background: Bone Marrow (BM) is an excellent source of Mesenchymal Stem Cells (MSCs) which can be expanded and used clinically. There is a growing evidence that MSCs can promote engraftment and reduce Graft versus Host Disease (GVHD). Bone marrow harvests are passed through 850μ, 500μ and 200μ filters to trap bony spicules and clots before infusion into patients. We hypothesize that the filters would also trap a significant number of MSCs which can be useful clinically after isolation and possible expansion. Methods: Bone marrow collection kits were obtained after informed consent of the donor (n = 11). Specimens were obtained in the course of scheduled bone marrow harvests. Cells were isolated and cultured from each portion of the device. Results: An average of 1×109 total mononuclear cells was obtained from the discarded marrow collection devices. Cells isolated in this experiment conform to published data regarding MSCs. The immunophenotype was CD44+, CD73+, CD90+, CD105+, CD14−, CD34−, CD45−, HLA-DR−, negative for leukocyte lineage specific antigens and positive for Prolyl-4-Hydroxylase. The cells were morphologically consistent with published data, being bilaterally symmetrical and forming conspicuous fibroblastoid colonies which readily grew to confluence and followed differentiation pathways to both adipose osteoid tissue. Fresh cells from the kits may also contain a precursor for MSCs that is CD90− and CD73−. Sorting for this precursor population yielded colony forming units consistent with MSCs. Conclusion: This represents a novel strategy for obtaining “healthy” material for potential therapy and study. No significant financial relationships to disclose.

scholarly journals The Immunoregulatory Effect of Mechanical Stress on Three-dimensional Cultured Mesenchymal Stem Cells After Extensive Expansion

2021 ◽  
Author(s):  
Fang-Ying Du ◽  
Na Zhao ◽  
Lei Bao ◽  
Jing Lei ◽  
An-Qi Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Mechanical Stress ◽  
Quantitative Polymerase Chain Reaction ◽  
Three Dimensional ◽  
3D Culture ◽  
Early Passage ◽  
Immunomodulatory Property ◽  
2D And 3D ◽  
Immunomodulatory Function

Abstract Background: Mesenchymal stem cells (MSCs) have been used to treat immunopathy, and three-dimensional (3D) cultured MSCs show enhanced immunomodulatory property compared with those in two-dimensional (2D) culture. However, both the regulatory mechanisms remain unclear. The aim of the study was to investigate the role of mechanical stress in maintaining the immunomodulatory function of 2D and 3D cultured MSCs.Methods: Umbilical cord mesenchymal stem cells (UC-MSCs) were plated on tissue culture plastic (TCP) as 2D culture and 3D cultured UC-MSCs were seeded in matrigel. Surface markers, clonogenicity, proliferation and immunoregulatory property of UC-MSCs were evaluated. Meanwhile, we established the mouse models of colitis and type 1 diabetes mellitus (T1DM) to reveal the pharmacotherapeutic effects of 3D cultured MSCs in vivo. The effect of changing mechanical stress by modulating Yes-associated protein (YAP) on immunomodulatory function of 2D and 3D cultured UC-MSCs was evaluated by immunofluorescent analysis, real-time quantitative polymerase chain reaction (qPCR) and western blot.Results: We verified early passage UC-MSCs in 2D and 3D cultures exhibited stemness, immunomodulatory property and therapeutic efficacy against immunopathy. However, these characteristics of 2D cultured UC-MSCs were impaired after extensive expansion, whereas 3D culture extended them for several passages by activating YAP. Moreover, prostaglandin E2 (PGE2) could up-regulate YAP to improve the immunomodulatory ability of 2D cultured UC-MSCs after extensive expansion. Conclusions: This work found for the first time that the significance of mechanical stress in maintaining immunoregulatory function of 2D and 3D cultured UC-MSCs, providing a new idea for improving the efficacy of MSCs-based immunotherapy.

scholarly journals Therapeutic Benefit for Late, but Not Early, Passage Mesenchymal Stem Cells on Pain Behaviour in an Animal Model of Osteoarthritis

2017 ◽  
Vol 2017 ◽  
pp. 1-11 ◽  
Author(s):  
Victoria Chapman ◽  
Hareklea Markides ◽  
Devi Rani Sagar ◽  
Luting Xu ◽  
James J. Burston ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Therapeutic Potential ◽  
Therapeutic Benefit ◽  
Therapeutic Effects ◽  
Early Passage ◽  
Pain Behaviour ◽  
Joint Pathology ◽  
Late Passage ◽  
Passage Number

Background. Mesenchymal stem cells (MSCs) have a therapeutic potential for the treatment of osteoarthritic (OA) joint pathology and pain. The aims of this study were to determine the influence of a passage number on the effects of MSCs on pain behaviour and cartilage and bone features in a rodent model of OA. Methods. Rats underwent either medial meniscal transection (MNX) or sham surgery under anaesthesia. Rats received intra-articular injection of either 1.5 × 106 late passage MSCs labelled with 10 μg/ml SiMAG, 1.5 × 106 late passage mesenchymal stem cells, the steroid Kenalog (200 μg/20 μL), 1.5 × 106 early passage MSCs, or serum-free media (SFM). Sham-operated rats received intra-articular injection of SFM. Pain behaviour was quantified until day 42 postmodel induction. Magnetic resonance imaging (MRI) was used to localise the labelled cells within the knee joint. Results. Late passage MSCs and Kenalog attenuated established pain behaviour in MNX rats, but did not alter MNX-induced joint pathology at the end of the study period. Early passage MSCs exacerbated MNX-induced pain behaviour for up to one week postinjection and did not alter joint pathology. Conclusion. Our data demonstrate for the first time the role of a passage number in influencing the therapeutic effects of MSCs in a model of OA pain.

scholarly journals Rejuvenated ageing mesenchymal stem cells by stepwise preconditioning ameliorates surgery-induced osteoarthritis in rabbits

2021 ◽  
Vol 10 (1) ◽  
pp. 10-21
Author(s):  
Zhixian Zong ◽  
Xiaoting Zhang ◽  
Zhengmeng Yang ◽  
Weihao Yuan ◽  
Jianping Huang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Therapeutic Effect ◽  
Subchondral Bone ◽  
Adult Stem Cells ◽  
Cruciate Ligament ◽  
Quantitative Polymerase Chain Reaction ◽  
Cartilage Damage ◽  
Human Mesenchymal Stem Cells ◽  
Early Passage

Aims Ageing-related incompetence becomes a major hurdle for the clinical translation of adult stem cells in the treatment of osteoarthritis (OA). This study aims to investigate the effect of stepwise preconditioning on cellular behaviours in human mesenchymal stem cells (hMSCs) from ageing patients, and to verify their therapeutic effect in an OA animal model. Methods Mesenchymal stem cells (MSCs) were isolated from ageing patients and preconditioned with chondrogenic differentiation medium, followed by normal growth medium. Cellular assays including Bromodeoxyuridine / 5-bromo-2'-deoxyuridine (BrdU), quantitative polymerase chain reaction (q-PCR), β-Gal, Rosette forming, and histological staining were compared in the manipulated human mesenchymal stem cells (hM-MSCs) and their controls. The anterior cruciate ligament transection (ACLT) rabbit models were locally injected with two millions, four millions, or eight millions of hM-MSCs or phosphate-buffered saline (PBS). Osteoarthritis Research Society International (OARSI) scoring was performed to measure the pathological changes in the affected joints after staining. Micro-CT analysis was conducted to determine the microstructural changes in subchondral bone. Results Stepwise preconditioning approach significantly enhanced the proliferation and chondrogenic potential of ageing hMSCs at early passage. Interestingly, remarkably lower immunogenicity and senescence was also found in hM-MSCs. Data from animal studies showed cartilage damage was retarded and subchondral bone remodelling was prevented by the treatment of preconditioned MSCs. The therapeutic effect depended on the number of cells applied to animals, with the best effect observed when treated with eight millions of hM-MSCs. Conclusion This study demonstrated a reliable and feasible stepwise preconditioning strategy to improve the safety and efficacy of ageing MSCs for the prevention of OA development. Cite this article: Bone Joint Res 2021;10(1):10–21.

Tripeptide-based macroporous hydrogel improves the osteogenic microenvironment of stem cells

2021 ◽  
Author(s):  
Qian Li ◽  
He Zhang ◽  
Jijia Pan ◽  
Binhong Teng ◽  
Ziqian Zeng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Three Dimensional ◽  
3D Culture ◽  
Human Mesenchymal Stem Cells ◽  
Osteogenic Induction

Due to the ability to combine multiple osteogenic induction "cues" at the same time, hydrogels are widely used in the three-dimensional (3D) culture of human mesenchymal stem cells (hMSCs) and...

Sign in / Sign up

Export Citation Format

Share Document