GENERATION OF 3-D PRINTED BONE TISSUE USING MESENCHYMAL STEM CELLS in vitro
SRINETHRA SRIDHAR, Dr. VIVIAN K. LEE SRM INSTITUTE OF SCIENCE AND TECHNOLOGY, BRIGHAM AND WOMEN’S HOSPITAL, HARVARD MEDICAL SCHOOL,MA,USA
ABSTRACT
OBJECTIVES
Perform differentiation assays Compare differentiated and undifferentiated cells Optimize concentration of components for 3 D culture Calculate cell viability Optimize differentiation of MSCs
METHODOLOGY
Growth of MSCS
Differentiatio n of MSCs
Assays to detect Differentiatio n of MSCs
3-D culture of bone cells
Cell counting and viability analysis
3D printing of bone tissue using bio printer
Bone tissue engineering is considered as one of the most promising field in the near future due to its huge contribution towards bone regeneration and development. Traditional bone repair methods like autografts, allografts though advantageous, prove to be less popular due to their major drawback: difficulties faced in harvest source tissues. Several other biomaterials like synthetic polymers, composite biomaterials have been developed over the past few years but these materials also have many disadvantages related to rate of disintegration, mechanical strength and biological safety. This project focuses on development of bone tissues using cellular method which is more biocompatible and biologically safe. Human mesenchymal stem cells from the bone marrow are used to differentiate into osteocytes and osteoblasts, the main bone forming cells using osteogenic differentiation media. The osteogenic media contains three main osteogenesis inducing components: Dexamethasone, Ascorbic Acid and β-glycerol phosphate. Mesenchymal stem cells are cultured using the osteogenic media in a 3-D environment in order to mimic in vivo environment of the body. To create an optimum 3-D biological environment, the cells are encapsulated in a synthetic UV cross linkable hydrogel called GelMA which acts as a biomaterial for the cells to adhere, spread and proliferate. Live dead staining is done to analyze the viability of the cells under in vitro conditions. Cell count for viability analysis is calculated using ImageJ software. GelMA being a versatile tool for 3-D bioprinting, is further used as a bio ink containing bone cells depending upon the viability for printing a biological in vitro bone tissue. CELL ISOLATION THROUGH BIOPSY FROM PATIENT GROWTH OF CELLS ON SCAFFOLD
INVITRO CELL CULTURE REGULATORY SIGNALS
3D BIOPRINTING
IMPLANTATION INTO THE PATIENT
TISSUE ENGINEERING PROCESS RESEARCH POSTER PRESENTATION DESIGN © 2015
www.PosterPresentations.com
RESULTS
DIFFERENTIATION OF MSCs
GELMA
GELMA + BM
MSC ENCAPSULATION
CELL VIABILITY PERCENTAGE
CONCLUSION
• Mesenchymal cells can be cultured and maintained to culture bone cells in 2D and 3D environment • Though GelMA is highly bio-compatible and helps in growth of cells in vitro, rheological properties need to be optimized. • In combination with highly viscous gels like alginate and carboxymethyl cellulose, GelMA can be used as a better bio ink. • MSC encapsulation can therefore be more successful in terms of cell viability. ACKNOWLEDGEMENT
I am thankful to SRMIST, Chennai for giving me the opportunity to do this research work under SAP PROGRAMME and my mentor Dr. Vivian K Lee who has provided all the support needed CHALLENGES IN TISSUE ENGINEERING to successfully complete the project.