Conversion of mouse embryonic fibroblast and bone marrow mesenchymal stem cells into functional osteoblast by defined factors

目录

Abstract

Contents

List of Tables

List of Figures

List of abbreviation

1．Bone biology

1．2 Bone anatomy

1．3 Bone components

1．4 Bone development(formation)

1．5 Bone remodeling

1．6 Bone healing

2．The Need for Bone Repair and Current Therapies

2．1 The need for bone repair

2．2 The current treatments for bone repair

3．Tissue engineering strategies for bone regeneration

3．1 Cell based strategies

3．2 Osteoinductive factors

3．3 Biomaterials(Scaffold materials)

4．Gene therapy strategies for bone regeneration

4．1 Methods of Gene delivery

4．2 Vectors for gene delivery

5．Lineage reprogramming

5．1 Trans-differentiation(trans-determination or direct conversion)

5．2 Direct reprogramming

References

Chapter 2．Conversion of mouse embryonic fibroblasts(MEF)into functional osteoblasts by defined factors

1．Introduction

2．Material and methods

2．1 Genetic materials

2．2 Cells

2．3 Study design

2．4In vitro osteogenic Induction

2．5 Characterization of the osteogenic induction

2．6 Statistical analysis

3．Results

3．1 PCR amplification of the interested factors

3．2 Confirmation of the successful cloning of the desired genes

3．3 Confirmation of the correct DNA Identity

3．4 Isolation and culture of MEF cells

3．5 Lentivirus transduction efficiency and the multiplicity of infection

3．6 Induction of osteoblasts from MEF by the combination of hLMP-3 and Yamanaka factors

3．7 Conversion efficiency of induced osteoblasts from c-Myc，Oct4 and hLMP-3 combination

3．8 MEF cells were directly reprogrammed into osteoblast like cells without passing through an intermediate pluripotency stage

4．Discussion

References

Chapter 3．In vivo stuay of the osteoblast formed cells after direct reprogramming of MEFs to form new bone in an induced bone defect

1．Introduction

2．Material and methods

2．1 Animal model

2．2 MEF cells isolation，transduction，and characterization

2．3 Cell seeding and scaffold material

2．4 Animal anesthesia and surgical procedures

2．5 Evaluation of the transplantation procedure

3．Results

3．1 Induction of a unilateral cortical femoral defect in SD rat

3．2 Osteoblasts from the COL group induced bone repair after transplantation into a unilateral bone defect

4．Discussion

References

Chapter 4．Comparison between curcumin and all-trans retinoic acid in the osteogenic differentiation of mouse bone marrow mesenchymal stem cells

1．Introduction

2．Material and methods

2．1 Experimental animals

2．2 Cells

2．3 Study design

2．4In vitro osteogenic Induction

2．5 Characterization of the osteogenic differentiation

2．6 Statistical analysis

3．Results

3．1 Morphological and immunotyping characterization of mouse BMSCs

3．2 Morphological changes during the osteogenic differentiation of BMSCs

3．3 Osteogenic differentiation capacity of BMSCs after induction with curcumin and ATRA

3．4 The effect of curcumin and ATRA on the expression level of the bone associated gene markers during BMSCs osteogenic differentiation

3．5 Expression of the OCN during the osteogenic differentiation process

3．6 Effect of curcumin supplemented OM on the osteogenic differentiation of lentivirus transduced MEFs with hLMP-3

4．Discussion

References

Conclusion

Innovations

Acknowledgements

Publications

声明