Polysaccharide Hydrogels Support the Long-Term Viability of Encapsulated Human Mesenchymal Stem Cells and Their Ability to Secrete Immunomodulatory Factors

While therapeutically interesting, the injection of MSCs suffers major limitations including cell death upon injection and a massive leakage outside the injection site. We proposed to entrap MSCs within spherical particles derived from alginate, as a control, or from silanized hydroxypropyl methylcellulose (Si-HPMC). We developed water in an oil dispersion method to produce small Si-HPMC particles with an average size of about 68 μm. We evidenced a faster diffusion of fluorescein isothiocyanate-dextran in Si-HPMC particles than in alginate ones. Human adipose-derived MSCs (hASC) were encapsulated either in alginate or in Si-HPMC, and the cellularized particles were cultured for up to 1 month. Both alginate and Si-HPMC particles supported cell survival, and the average number of encapsulated hASC per alginate and Si-HPMC particle (7102 and 5100, resp.) did not significantly change. The stimulation of encapsulated hASC with proinflammatory cytokines resulted in the production of IDO, PGE2, and HGF whose concentration was always higher when cells were encapsulated in Si-HPMC particles than in alginate ones. We have demonstrated that Si-HPMC and alginate particles support hASC viability and the maintenance of their ability to secrete therapeutic factors.

Download Full-text

In Vitro Assessment of Hydroxyapatite Coating on the Surface of Additive Manufactured Ti6Al4V Scaffolds

Materials Science Forum ◽

10.4028/www.scientific.net/msf.879.2444 ◽

2016 ◽

Vol 879 ◽

pp. 2444-2449 ◽

Cited By ~ 1

Author(s):

Ekaterina Chudinova ◽

Maria Surmeneva ◽

Andrey Koptioug ◽

Irina V. Savintseva ◽

Irina Selezneva ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Human Mesenchymal Stem Cells ◽

Nanocrystalline Structure ◽

X Ray Diffraction ◽

Ha Coating ◽

Magnetron Sputter Deposition ◽

In Vitro Assessment ◽

Average Size

Custom orthopedic and dental implants may be fabricated by additive manufacturing (AM), for example using electron beam melting technology. This study is focused on the modification of the surface of Ti6Al4V alloy coin-like scaffolds fabricated via AM technology (EBM®) by radio frequency (RF) magnetron sputter deposition of hydroxyapatite (HA) coating. The scaffolds with HA coating were characterized by Scanning Electron microscopy, X-ray diffraction. HA coating showed a nanocrystalline structure with the crystallites of an average size of 32±9 nm. The ability of the surface to support adhesion and the proliferation of human mesenchymal stem cells was studied using biological short-term tests in vitro. In according to in vitro assessment, thin HA coating stimulated the attachment and proliferation of cells. Human mesenchymal stem cells cultured on the HA-coated scaffold also formed mineralized nodules.

Download Full-text

MYOGENIC DIFFERENTIATION OF HUMAN MESENCHYMAL STEM CELLS IN VITRO AND LONG-TERM SURVIVAL IN A NUDE RAT MODEL FOR URINARY INCONTINENCE

The Journal of Urology ◽

10.1016/s0022-5347(09)61907-8 ◽

2009 ◽

Vol 181 (4) ◽

pp. 681

Author(s):

Gerhard Feil ◽

Adriana Drost ◽

Simon Baumann ◽

Jochen Schäfer ◽

Sibylle Weng ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Urinary Incontinence ◽

Rat Model ◽

Myogenic Differentiation ◽

Human Mesenchymal Stem Cells ◽

Term Survival ◽

Nude Rat

Download Full-text

Soft Substrate Maintains Proliferative and Adipogenic Differentiation Potential of human Mesenchymal Stem Cells on Long Term Expansion by Delaying Senescence

10.1101/364059 ◽

2018 ◽

Author(s):

Sanjay K. Kureel ◽

Pankaj Mogha ◽

Akshada Khadpekar ◽

Vardhman Kumar ◽

Rohit Joshi ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Culture System ◽

Human Mesenchymal Stem Cells ◽

Population Doubling ◽

Differentiation Potential ◽

Lineage Differentiation ◽

Poly Acrylamide

AbstractHuman mesenchymal stem cells (hMSCs), when cultured on tissue culture plate (TCP) for in vitro expansion, they spontaneously lose their proliferative capacity and multi-lineage differentiation potential. They also lose their distinct spindle morphology and become large and flat. After a certain number of population doubling, they enter into permanent cell cycle arrest, called senescence. This is a major roadblock for clinical use of hMSCs which demands large number of cells. A cell culture system is needed which can maintain the stemness of hMSCs over long term passages yet simple to use. In this study, we explore the role of substrate rigidity in maintaining stemness. hMSCs were serially passaged on TCP and 5 kPa poly-acrylamide gel for 20 population doubling. It was found that while on TCP, cell growth reached a plateau at cumulative population doubling (CPD) = 12.5, on 5 kPa gel, they continue to proliferate linearly till we monitored (CPD = 20). We also found that while on TCP, late passage MSCs lost their adipogenic potential, the same was maintained on soft gel. Cell surface markers related to MSCs were also unaltered. We demonstrated that this maintenance of stemness was correlated with delay in onset of senescence, which was confirmed by β-gal assay and by differential expression of vimentin, Lamin A and Lamin B. As preparation of poly-acrylamide gel is a simple, well established, and well standardized protocol, we believe that this system of cell expansion will be useful in therapeutic and research applications of hMSCs.One Sentence SummaryhMSCs retain their stemness when expanded in vitro on soft polyacrylamide gel coated with collagen by delaying senescence.Significance StatementFor clinical applications, mesenchymal stem cells (MSCs) are required in large numbers. As MSCs are available only in scarcity in vivo, to fulfill the need, extensive in vitro expansion is unavoidable. However, on expansion, they lose their replicative and multi-lineage differentiation potential and become senescent. A culture system that can maintain MSC stemness on long-term expansion, without compromising the stemness, is need of the hour. In this paper, we identified polyacrylamide (PAA) hydrogel of optimum stiffness that can be used to maintain stemness of MSCs during in vitro long term culture. Large quantity of MSCs thus grown can be used in regenerative medicine, cell therapy, and in treatment of inflammatory diseases.

Download Full-text

A Decrease in the Proliferative Activity of Human Mesenchymal Stem Cells during Long-Term Cultivation is Not Connected with Change in Their Migration Properties

Cell and Tissue Biology ◽

10.1134/s1990519x18030070 ◽

2018 ◽

Vol 12 (3) ◽

pp. 197-206 ◽

Cited By ~ 1

Author(s):

N. F. Mikheeva ◽

P. A. Butylin ◽

A. Yu. Zaritskii ◽

B. V. Popov

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Proliferative Activity ◽

Human Mesenchymal Stem Cells

Download Full-text

Long-Term Quantitative Biodistribution and Side Effects of Human Mesenchymal Stem Cells (hMSCs) Engraftment in NOD/SCID Mice following Irradiation

Stem Cells International ◽

10.1155/2014/939275 ◽

2014 ◽

Vol 2014 ◽

pp. 1-13 ◽

Cited By ~ 7

Author(s):

Sabine François ◽

Benoit Usunier ◽

Luc Douay ◽

Marc Benderitter ◽

Alain Chapel

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Side Effects ◽

Total Body Irradiation ◽

Human Mesenchymal Stem Cells ◽

Scid Mice ◽

Human Mscs ◽

Long Term Treatment ◽

Total Body

There is little information on the fate of infused mesenchymal stem cells (MSCs) and long-term side effects after irradiation exposure. We addressed these questions using human MSCs (hMSCs) intravenously infused to nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice submitted to total body irradiation (TBI) or local irradiation (abdominal or leg irradiation). The animals were sacrificed 3 to 120 days after irradiation and the quantitative and spatial distribution of hMSCs were studied by polymerase chain reaction (PCR). Following their infusion into nonirradiated animals, hMSCs homed to various tissues. Engraftment depended on the dose of irradiation and the area exposed. Total body irradiation induced an increased hMSC engraftment level compared to nonirradiated mice, while local irradiations increased hMSC engraftment locally in the area of irradiation. Long-term engraftment of systemically administered hMSCs in NOD/SCID mice increased significantly in response to tissue injuries produced by local or total body irradiation until 2 weeks then slowly decreased depending on organs and the configuration of irradiation. In all cases, no tissue abnormality or abnormal hMSCs proliferation was observed at 120 days after irradiation. This work supports the safe and efficient use of MSCs by injection as an alternative approach in the short- and long-term treatment of severe complications after radiotherapy for patients refractory to conventional treatments.

Download Full-text