Kentaro Onishi1, Dakota L Jones2, Scott M Riester3, Eric A Lewallen4, David G Lewallen5, Jacob L Sellon6, Allan B Dietz7, Wenchun Qu8, Andre J van Wijnen9, Jay Smith10. 1. Department of Physical Medicine & Rehabilitation, Mayo Clinic Sports Medicine Center, Mayo Clinic, Rochester, MN(∗). 2. Department of Biomedical Engineering and Physiology, Mayo Graduate School, Mayo Clinic, Rochester, MN; Department of Orthopedic Surgery, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN(†). 3. Department of Orthopedic Surgery, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN(‡). 4. Department of Orthopedic Surgery, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN(§). 5. Department of Orthopedic Surgery, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN(‖). 6. Department of Physical Medicine & Rehabilitation, Mayo Clinic Sports Medicine Center, Mayo Clinic, Rochester, MN(¶). 7. Department of Biochemistry & Molecular Biology, Mayo Graduate School, Mayo Clinic, Rochester, MN; Department of Laboratory Medicine & Pathology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN(#). 8. Department of Physical Medicine & Rehabilitation, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN; Department of Anesthesiology Division of Pain Medicine, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN(∗∗). 9. Department of Orthopedic Surgery, Medical Sciences Building, Rm S3-69, Mayo Clinic, 200 1st St, SW, Rochester, MN 55905; Department of Biomedical Engineering and Physiology, Mayo Graduate School, Mayo Clinic, Rochester, MN; Department of Biochemistry & Molecular Biology, Mayo Graduate School, Mayo Clinic, Rochester, MN(††). Electronic address: vanwijnen.andre@mayo.edu. 10. Department of Physical Medicine & Rehabilitation, W14, Mayo Building, Mayo Clinic, 200 1st St, SW, Rochester, MN 55905; Department of Radiology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN; Department of Anatomy, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN(‡‡). Electronic address: Smith.jay@mayo.edu.
Abstract
OBJECTIVE: To assess the biological effects of passage through clinically relevant needles on the viability and metabolic activity of culture-expanded, human adipose tissue-derived mesenchymal stromal/stem cells (AMSCs). DESIGN: Prospective observational pilot study. SETTING: Academic medical center. PARTICIPANTS: Patient-derived clinical-grade culture expanded AMSCs. INTERVENTIONS: AMSCs were passed through syringes without a needle attached (control), with an 18-gauge (25.4-mm) needle attached and with a 30-gauge (19-mm) needle attached at a constant injection flow rate and constant cell concentrations. Each injection condition was completed in triplicate. MAIN OUTCOME MEASURES: Cell number and viability, proliferative capacity, metabolic activity, and acute gene expression as measured by cell counts, mitochondrial activity, and quantitative real time reverse-transcription polymerase chain reaction on day 0 (immediately), day 1, and day 4 after injection. RESULTS: AMSC viability was not significantly affected by injection, and cells proliferated normally regardless of study group. Postinjection, AMSCs robustly expressed both proliferation markers and extracellular matrix proteins. Stress-response mRNAs were markedly but transiently increased independently of needle size within the first day in culture postinjection. CONCLUSIONS: Human, culture-expanded AMSCs maintain their viability, proliferative capacity, and metabolic function following passage through needles as small as 30-gauge at constant flow rates of 4 mL/min, despite an early, nonspecific stress/cytoprotective response. These initial findings suggest that culture-expanded AMSCs should tolerate the injection process during most cell-based therapeutic interventions.
OBJECTIVE: To assess the biological effects of passage through clinically relevant needles on the viability and metabolic activity of culture-expanded, human adipose tissue-derived mesenchymal stromal/stem cells (AMSCs). DESIGN: Prospective observational pilot study. SETTING: Academic medical center. PARTICIPANTS: Patient-derived clinical-grade culture expanded AMSCs. INTERVENTIONS: AMSCs were passed through syringes without a needle attached (control), with an 18-gauge (25.4-mm) needle attached and with a 30-gauge (19-mm) needle attached at a constant injection flow rate and constant cell concentrations. Each injection condition was completed in triplicate. MAIN OUTCOME MEASURES: Cell number and viability, proliferative capacity, metabolic activity, and acute gene expression as measured by cell counts, mitochondrial activity, and quantitative real time reverse-transcription polymerase chain reaction on day 0 (immediately), day 1, and day 4 after injection. RESULTS: AMSC viability was not significantly affected by injection, and cells proliferated normally regardless of study group. Postinjection, AMSCs robustly expressed both proliferation markers and extracellular matrix proteins. Stress-response mRNAs were markedly but transiently increased independently of needle size within the first day in culture postinjection. CONCLUSIONS:Human, culture-expanded AMSCs maintain their viability, proliferative capacity, and metabolic function following passage through needles as small as 30-gauge at constant flow rates of 4 mL/min, despite an early, nonspecific stress/cytoprotective response. These initial findings suggest that culture-expanded AMSCs should tolerate the injection process during most cell-based therapeutic interventions.
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