The Combined Thermoresponsive Cell-Imprinted Substrate, Induced Differentiation, and "KLC Sheet" Formation

Purpose: Stem cells can exhibit restorative effects with the commitment to functional cells. Cell-imprinted topographies provide adaptable templates and certain dimensions for the differentiation and bioactivity of stem cells. Cell sheet technology using the thermo-responsive polymers detaches the "cell sheets" easier with less destructive effects on the extracellular matrix (ECM). Here, we aim to dictate keratinocyte-like differentiation of mesenchymal stem cells by using combined cell imprinting and sheet technology. Methods: We developed the poly dimethyl siloxane (PDMS) substrate having keratinocyte cell-imprinted topography grafted with the PNIPAAm polymer. Adipose tissue-derived mesenchymal stem cells (AT-MSCs) were cultured on PDMS substrate for 14 days and keratinocyte-like differentiation monitored via the expression of involucrin, P63, and cytokeratin 14. Results: Data showed the efficiency of the current protocol in the fabrication of PDMS molds. The culture of AT-MSCs induced typical keratinocyte morphology and up-regulated the expression of cytokeratin-14, Involucrin, and P63 compared to AT-MSCs cultured on the plastic surface (p<0.05). Besides, KLC sheets were generated once slight changes occur in the environment temperature. Conclusion: These data showed the hypothesis that keratinocyte cell imprinted substrate can orient AT-MSCs toward KLCs by providing a specific niche and topography.

Download Full-text

Self-assembled cell sheets composed of mesenchymal stem cells and gelatin nanofibers for the treatment of full-thickness wounds

Biomaterials Science ◽

10.1039/d0bm00910e ◽

2020 ◽

Vol 8 (16) ◽

pp. 4535-4544

Author(s):

Oanh-Vu Pham-Nguyen ◽

Ji Un Shin ◽

Hyesung Kim ◽

Hyuk Sang Yoo

Keyword(s):

Stem Cells ◽

Wound Healing ◽

Mesenchymal Stem Cells ◽

Cell Sheet ◽

Full Thickness ◽

Cell Sheets ◽

Self Assembled

Gelatin-layered PCL nanofibrils for 3D cell sheet formation were composed with adipocyte-derived stem cells for wound healing.

Download Full-text

Effect of Gelatin on Osteogenic Cell Sheet Formation Using Canine Adipose-Derived Mesenchymal Stem Cells

Cell Transplantation ◽

10.3727/096368916x693338 ◽

2017 ◽

Vol 26 (1) ◽

pp. 115-123 ◽

Cited By ~ 15

Author(s):

Ah Young Kim ◽

Yongsun Kim ◽

Seung Hoon Lee ◽

Yongseok Yoon ◽

Wan-Hee Kim ◽

...

Keyword(s):

Stem Cells ◽

Cell Proliferation ◽

Mesenchymal Stem Cells ◽

Bone Regeneration ◽

Cell Sheet ◽

Proliferation Rate ◽

Osteogenic Cell ◽

Cell Sheets ◽

Gene Markers ◽

Cell Proliferation Rate

Osteogenically differentiated cell sheet techniques using mesenchymal stem cells (MSCs) are available to stimulate bone regeneration. The advantage of the cell sheet technique is delivering live cells effectively into the focal region. We developed a novel osteogenic cell sheet technique by adding gelatin to osteogenic cell medium. Gelatin-induced osteogenic cell sheets (GCSs) were compared to conventional osteogenic cell sheets (OCSs). Undifferentiated MSCs (UCs) were used as a control. The morphology of these cell sheets was evaluated microscopically and histologically. The time-dependent cell proliferation rate was estimated by DNA quantification. The expression of osteogenic gene markers and the number of calcium depositions were assessed by quantitative real-time polymerase chain reaction and Alizarin red S (ARS) staining, respectively. GCSs were thicker and stronger than OCSs. GCSs showed a significantly higher cell proliferation rate compared to OCSs ( p < 0.05). GCSs exhibited significantly higher upregulation of BMP-7 mRNA compared to OCSs ( p < 0.05). Both GCSs and OCSs showed negative ARS reactivity on day 10, but only GCSs showed positive ARS reactivity on day 21. With this technique, we observed active cell proliferation with abundant ECM and upregulation of osteogenic bone markers, and our results suggest that GCSs could be promising for therapeutic applications in bone regeneration.

Download Full-text

Decellularized Articular Cartilage Microparticles for Expansion of Mesenchymal Stem Cells and Zonal Regeneration of Articular Cartilage

10.1101/2021.04.16.440121 ◽

2021 ◽

Author(s):

Azadeh Sepahvandi ◽

Safaa Ibrahim Kader ◽

Victor Anthony Madormo ◽

Mehri Monavarian ◽

Esmaiel Jabbari

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Articular Cartilage ◽

Cell Sheet ◽

Compressive Modulus ◽

Human Mscs ◽

Cell Sheets ◽

Chondrogenic Medium ◽

Alginate Gel ◽

Stratified Structure

Introduction: The objective was to create multilayer cellular constructs using fetal or adult, decellularized articular cartilage in particulate form as microcarriers for expansion and fusion of mesenchymal stem cells (MSCs) to regenerate the stratified structure of articular cartilage. Methods: Porous microparticles (CMPs) generated from decellularized fetal or adult bovine articular cartilage were used as microcarriers for expansion of human MSCs. The CMP expanded MSCs (CMP-MSCs) were used to generate injectable hydrogels or preformed multilayer constructs for articular cartilage regeneration. In the injectable approach, CMP-MSCs were suspended in alginate gel, crosslinked with calcium chloride, and incubated in chondrogenic medium to generate an injectable regenerative construct. In the preformed approach, fetal or adult CMP-MSCs were suspended in a culture medium, allowed to settle sequentially by the force of gravity, and fused by incubation in chondrogenic medium to generate multilayer cell sheets. The constructs were characterized with respect to compressive modulus, cellularity, and expression of chondrogenic markers. Results: Human MSCs expanded on fetal or adult CMPs in basal medium maintained the expression of mesenchymal markers. The injectable CMP-MSCs hydrogels had significantly higher expression of chondrogenic markers and compressive modulus after four weeks incubation in chondrogenic medium compared to MSCs directly encapsulated in alginate gel; preformed CMP-MSCs cell sheets had significantly higher compressive modulus and expression of chondrogenic markers compared to MSCs in the pellet culture. Conclusion: The preformed cell sheet approach is potentially useful for creating multilayer constructs by sequential gravitational settling of CMP-MSCs to mimic the stratified structure of articular cartilage.

Download Full-text

Cell sheet formation enhances the therapeutic effects of human umbilical cord mesenchymal stem cells on myocardial infarction as a bioactive material

Bioactive Materials ◽

10.1016/j.bioactmat.2021.01.036 ◽

2021 ◽

Vol 6 (9) ◽

pp. 2999-3012

Author(s):

Rui Guo ◽

Feng Wan ◽

Masatoshi Morimatsu ◽

Qing Xu ◽

Tian Feng ◽

...

Keyword(s):

Myocardial Infarction ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Umbilical Cord ◽

Cell Sheet ◽

Human Umbilical Cord ◽

Therapeutic Effects ◽

Bioactive Material

Download Full-text

Shrinkage-free preparation of scaffold-free cartilage-like disk-shaped cell sheet using human bone marrow mesenchymal stem cells

Journal of Bioscience and Bioengineering ◽

10.1016/j.jbiosc.2010.11.022 ◽

2011 ◽

Vol 111 (4) ◽

pp. 489-492 ◽

Cited By ~ 13

Author(s):

Satoshi Maeda ◽

Takashi Fujitomo ◽

Takahiro Okabe ◽

Shigeyuki Wakitani ◽

Mutsumi Takagi

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Cell Sheet ◽

Human Bone ◽

Human Bone Marrow ◽

Shaped Cell ◽

Marrow Mesenchymal Stem Cells

Download Full-text

Hepatic cell sheets engineered from human mesenchymal stem cells with a single small molecule compound IC-2 ameliorate acute liver injury in mice

Regenerative Therapy ◽

10.1016/j.reth.2018.07.001 ◽

2018 ◽

Vol 9 ◽

pp. 45-57 ◽

Cited By ~ 5

Author(s):

Noriko Itaba ◽

Ikuya Noda ◽

Hiroyuki Oka ◽

Yohei Kono ◽

Kaori Okinaka ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Liver Injury ◽

Small Molecule ◽

Acute Liver Injury ◽

Human Mesenchymal Stem Cells ◽

Hepatic Cell ◽

Cell Sheets ◽

Small Molecule Compound

Download Full-text

Mechanical stress potentiates the differentiation of periodontal ligament stem cells into keratocytes

British Journal of Ophthalmology ◽

10.1136/bjophthalmol-2017-311150 ◽

2018 ◽

Vol 102 (4) ◽

pp. 562-569 ◽

Cited By ~ 5

Author(s):

Jialin Chen ◽

Wei Zhang ◽

Ludvig J Backman ◽

Peyman Kelk ◽

Patrik Danielson

Keyword(s):

Stem Cells ◽

Mechanical Stimulation ◽

Periodontal Ligament ◽

Synergistic Effects ◽

Mechanical Strain ◽

Cell Sheet ◽

Corneal Stroma ◽

Cell Sheets ◽

Periodontal Ligament Stem Cells ◽

Gene And Protein Expression

AimsTo explore the role of corneal-shaped static mechanical strain on the differentiation of human periodontal ligament stem cells (PDLSCs) into keratocytes and the possible synergistic effects of mechanics and inducing medium.MethodsPDLSCs were exposed to 3% static dome-shaped mechanical strain in a Flexcell Tension System for 3 days and 7 days. Keratocyte phenotype was determined by gene expression of keratocyte markers. Keratocyte differentiation (inducing) medium was introduced in the Flexcell system, either continuously or intermittently combined with mechanical stimulation. The synergistic effects of mechanics and inducing medium on keratocyte differentiation was evaluated by gene and protein expression of keratocyte markers. Finally, a multilamellar cell sheet was assembled by seeding PDLSCs on a collagen membrane and inducing keratocyte differentiation. The transparency of the cell sheet was assessed, and typical markers of native human corneal stroma were evaluated by immunofluorescence staining.ResultsDome-shaped mechanical stimulation promoted PDLSCs to differentiate into keratocytes, as shown by the upregulation of ALDH3A1, CD34, LUM, COL I and COL V. The expression of integrins were also upregulated after mechanical stimulation, including integrin alpha 1, alpha 2, beta 1 and non-muscle myosin II B. A synergistic effect of mechanics and inducing medium was found on keratocyte differentiation. The cell sheets were assembled under the treatment of mechanics and inducing medium simultaneously. The cell sheets were transparent, multilamellar and expressed typical markers of corneal stroma.ConclusionDome-shaped mechanical stimulation promotes differentiation of PDLSCs into keratocytes and has synergistic effects with inducing medium. Multilamellar cell sheets that resemble native human corneal stroma show potential for future clinical applications.

Download Full-text

Characteristic differences of cell sheets composed of mesenchymal stem cells with different tissue origins

Regenerative Therapy ◽

10.1016/j.reth.2019.01.002 ◽

2019 ◽

Vol 11 ◽

pp. 34-40 ◽

Cited By ~ 4

Author(s):

Mitsuyoshi Nakao ◽

Daimu Inanaga ◽

Kenichi Nagase ◽

Hideko Kanazawa

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Cell Sheets

Download Full-text

A Concise Review on the Use of Mesenchymal Stem Cells in Cell Sheet-Based Tissue Engineering with Special Emphasis on Bone Tissue Regeneration

Stem Cells International ◽

10.1155/2017/2374161 ◽

2017 ◽

Vol 2017 ◽

pp. 1-13 ◽

Cited By ~ 27

Author(s):

A. Cagdas Yorukoglu ◽

A. Esat Kiter ◽

Semih Akkaya ◽

N. Lale Satiroglu-Tufan ◽

A. Cevik Tufan

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Ethical Issues ◽

Cell Sheet ◽

Bone Tissue Regeneration ◽

Differentiation Potential ◽

Cell Sheet Engineering ◽

Application Techniques

The integration of stem cell technology and cell sheet engineering improved the potential use of cell sheet products in regenerative medicine. This review will discuss the use of mesenchymal stem cells (MSCs) in cell sheet-based tissue engineering. Besides their adhesiveness to plastic surfaces and their extensive differentiation potential in vitro, MSCs are easily accessible, expandable in vitro with acceptable genomic stability, and few ethical issues. With all these advantages, they are extremely well suited for cell sheet-based tissue engineering. This review will focus on the use of MSC sheets in osteogenic tissue engineering. Potential application techniques with or without scaffolds and/or grafts will be discussed. Finally, the importance of osteogenic induction of these MSC sheets in orthopaedic applications will be demonstrated.

Download Full-text