Adipogenic differentiation of laser-printed 3D tissue grafts consisting of human adipose-derived stem cells

Laser-assisted bioprinting (LaBP) allows the realization of computer-generated 3D tissue grafts consisting of cells embedded in a hydrogel environment. In this study, human adipose-derived stem cells (hASCs) were printed in a free-scalable 3D grid pattern by means of LaBP. We demonstrate that neither the proliferation ability nor the differentiation behaviour of the stem cells was affected by the LaBP procedure. Furthermore, the 3D grafts were differentiated down the adipogenic lineage pathway for 10 days. We verify by quantitative assessments of adipogenic markers that the 3D grafts resemble cell lineages present in natural adipose tissue. Additionally, we provide the proof that even pre-differentiated hASCs could be utilized for the generation of 3D tissue grafts. These results indicate that the biofabrication of living grafts resembling their complex native origin is within reach.

[1]  Birgit Glasmacher,et al.  Laser printing of stem cells for biofabrication of scaffold-free autologous grafts. , 2011, Tissue engineering. Part C, Methods.

[2]  Ian M. Hutchings,et al.  Direct Writing Technology Advances and Developments , 2008 .

[3]  Nan Ma,et al.  Laser printing of skin cells and human stem cells. , 2010, Tissue engineering. Part C, Methods.

[4]  C T Laurencin,et al.  Extracellular matrix production by human osteoblasts cultured on biodegradable polymers applicable for tissue engineering. , 2003, Biomaterials.

[5]  Bradley R. Ringeisen,et al.  Laser Printing of Single Cells: Statistical Analysis, Cell Viability, and Stress , 2005, Annals of Biomedical Engineering.

[6]  Yong Huang,et al.  Laser-based direct-write techniques for cell printing , 2010, Biofabrication.

[7]  R. Hynes,et al.  Ras Activation Is Necessary for Integrin-mediated Activation of Extracellular Signal-regulated Kinase 2 and Cytosolic Phospholipase A2 but Not for Cytoskeletal Organization* , 1996, The Journal of Biological Chemistry.

[8]  N. Cooper,et al.  β1 integrin antagonism on adherent, differentiated human neuroblastoma cells triggers an apoptotic signaling pathway , 2000, Neuroscience.

[9]  Bradley R Ringeisen,et al.  Single-cell printing to form three-dimensional lines of olfactory ensheathing cells , 2008, Biomedical materials.

[10]  R. Zhao,et al.  Bone Marrow-Derived Stem / Progenitor Cells in Cutaneous Repair and Regeneration , 2010 .

[11]  M. Hedrick,et al.  Fat tissue: an underappreciated source of stem cells for biotechnology. , 2006, Trends in biotechnology.

[12]  Nils Cordes,et al.  Signalling via integrins: implications for cell survival and anticancer strategies. , 2007, Biochimica et biophysica acta.

[13]  J. Lannutti,et al.  Nanotopographic control of cytoskeletal organization. , 2006, Langmuir.

[14]  R L Juliano,et al.  Cell adhesion molecules, signal transduction and cell growth. , 1999, Current opinion in cell biology.

[15]  J. M. Fernández-Pradas,et al.  Printing biological solutions through laser-induced forward transfer , 2008 .

[16]  Robert DiRaddo,et al.  Design and Dynamic Culture of 3D-Scaffolds for Cartilage Tissue Engineering , 2011, Journal of biomaterials applications.

[17]  Joshua C. Hansen,et al.  The regulation of integrin-mediated osteoblast focal adhesion and focal adhesion kinase expression by nanoscale topography. , 2007, Biomaterials.

[18]  A. Ngezahayo,et al.  Gap Junction Coupling and Apoptosis in GFSHR-17 Granulosa Cells , 2005, The Journal of Membrane Biology.

[19]  Juergen Koch,et al.  Time-resolved imaging of hydrogel printing via laser-induced forward transfer , 2011 .

[20]  L. Koch,et al.  Laser printing of cells into 3D scaffolds , 2010, Biofabrication.

[21]  N Lenoir,et al.  Europe confronts the embryonic stem cell research challenge. , 2000, Science.

[22]  A. Atala,et al.  Sources of Stem Cells for Regenerative Medicine , 2008, Stem Cell Reviews.

[23]  Bradley R Ringeisen,et al.  Jet‐based methods to print living cells , 2006, Biotechnology journal.

[24]  F. Guillemot,et al.  Laser assisted bioprinting of engineered tissue with high cell density and microscale organization. , 2010, Biomaterials.

[25]  Yong Huang,et al.  Droplet formation in matrix-assisted pulsed-laser evaporation direct writing of glycerol-water solution , 2009 .

[26]  K. Hochedlinger,et al.  Induced pluripotency: history, mechanisms, and applications. , 2010, Genes & development.

[27]  James J. Yoo,et al.  Tissue-engineered autologous bladders for patients needing cystoplasty , 2006, The Lancet.

[28]  A. Galinier,et al.  Adipose tissues as an ancestral immune organ: Site‐specific change in obesity , 2005, FEBS letters.

[29]  Zsolt Bor,et al.  Survival and proliferative ability of various living cell types after laser-induced forward transfer. , 2005, Tissue engineering.

[30]  Douglas B. Chrisey,et al.  The Power of Direct Writing , 2000, Science.

[31]  L. Silengo,et al.  Soluble integrin ligands and growth factors independently rescue neuroblastoma cells from apoptosis under nonadherent conditions. , 1997, Experimental cell research.

[32]  A. Bader,et al.  Multilineage differentiation potential of human dermal skin‐derived fibroblasts , 2008, Experimental dermatology.

[33]  L. Tartaglia,et al.  Chronic inflammation in fat plays a crucial role in the development of obesity-related insulin resistance. , 2003, The Journal of clinical investigation.

[34]  M. Desai,et al.  Obesity is associated with macrophage accumulation in adipose tissue. , 2003, The Journal of clinical investigation.

[35]  Xiaofeng Cui,et al.  Application of inkjet printing to tissue engineering , 2006, Biotechnology journal.

[36]  G. Vunjak‐Novakovic,et al.  Hypoxia and stem cell‐based engineering of mesenchymal tissues , 2009, Biotechnology progress.

[37]  Andrés J. García Get a grip: integrins in cell-biomaterial interactions. , 2005, Biomaterials.

[38]  H. Mizuno Adipose-derived stem cells for tissue repair and regeneration: ten years of research and a literature review. , 2009, Journal of Nippon Medical School = Nippon Ika Daigaku zasshi.

[39]  K. Tobita,et al.  Directed Stem Cell Differentiation: The Role of Physical Forces , 2010, Cell communication & adhesion.

[40]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.