1. ABSTRACT
2013 Summer Research Program
Office of Educational Programs
Medical School
Undergraduate Student
Isolation and Culture of Mesenchymal Stem Cells from Mouse Bone
Marrow
OSAMA A. JEBREEN Houston Baptist University Class of 2015
Sponsored by: Pauline J. Duke, PhD, Orthodontics
Supported by: : UTSD Research Office, Texas Space Grant Consortium
Key Words: Bone marrow stem cells, Isolation, Culturing, Lasering
Background: Bone-forming cartilage cells derived from mouse bone marrow stem cells (BMSC)
provide researchers with a method that can be applied in tissue engineering. Such an approach
is used to grow and form cartilage cells that can be beneficial in clinical use.
Objective: The objective was to isolate and culture mouse bone marrow stem cells into cartilage
cells with the assistance of low level laser therapy (LLT) to help reduce the amount of time that
cells need to grow and differentiate.
Materials&Methods: Femurs from four C57 black adult male mice were dissected away from
the mouse skeleton and rinsed in Phosphate Buffered Saline (PBS). Both ends of the femur were
removed, and a 27 Gauge (27G) needle attached to a 1-ml syringe containing BMSC complete
medium was inserted through the cut end of the femur to flush out the stem cells into T-25Cm2
flask. Cells in flasks were incubated under standard conditions (37°C, humidity, 5% Co2). 50%
of the medium was removed and replaced every two-three days. Floaters and loose cells were
removed and placed in a T-150Cm2
flask. Cells were attached after 24-48 hours and reached a
95% confluency. After 7 days, cells were trypsinized to a larger Flask (T-75Cm2
using 0.05%
Trypsin-EDTA). Cells were placed in the incubator with constantly changing the media for 7
days before counting them.
Results & Conclusion: We were only able to culture stem cells without the process of lasering
due to insufficient growth of cells.
Acknowledgments: Dr. Zhang, Dr. Cai, Dr. Barros, Dr. Patel, Dr. Tribble’s lab, Dina Montufar-
Solis, Adriana Cavender.