Clinical parameters affecting multipotent adult progenitor cells in vitro
Margaret L. Jackson,
Katherine A. Ruppert,
Daniel J. Kota,
Karthik S. Prabhakara,
Robert A. Hetz,
Benjamin M. Aertker,
Supinder Bedi,
Robert W. Mays,
Scott D. Olson,
Charles S. Cox, Jr.
Affiliations
Margaret L. Jackson
Departments of Pediatric Surgery, McGovern Medical School at the University of Texas Health Science Center at Houston, United States
Katherine A. Ruppert
Departments of Pediatric Surgery, McGovern Medical School at the University of Texas Health Science Center at Houston, United States
Daniel J. Kota
Emory Personalized Immunotherapy Core, Emory University, United States
Karthik S. Prabhakara
Departments of Pediatric Surgery, McGovern Medical School at the University of Texas Health Science Center at Houston, United States
Robert A. Hetz
Departments of Pediatric Surgery, McGovern Medical School at the University of Texas Health Science Center at Houston, United States; Departments of Surgery, McGovern Medical School at the University of Texas Health Science Center at Houston, United States
Benjamin M. Aertker
Departments of Pediatric Surgery, McGovern Medical School at the University of Texas Health Science Center at Houston, United States
Supinder Bedi
Departments of Pediatric Surgery, McGovern Medical School at the University of Texas Health Science Center at Houston, United States
Robert W. Mays
Athersys, Inc, United States
Scott D. Olson
Departments of Pediatric Surgery, McGovern Medical School at the University of Texas Health Science Center at Houston, United States; Corresponding authors.
Charles S. Cox, Jr.
Departments of Pediatric Surgery, McGovern Medical School at the University of Texas Health Science Center at Houston, United States; Departments of Surgery, McGovern Medical School at the University of Texas Health Science Center at Houston, United States; Corresponding authors.
Background: Human multipotent adult progenitor cells (MAPC®) are an emerging therapy for traumatic brain injury (TBI); however, clinically translating a therapy involves overcoming many factors in vivo which are not present in pre-clinical testing. In this study we examined clinical parameters in vitro that may impact cell therapy efficacy. Methods: MAPC were infused through varying gauged needles and catheters with and without chlorhexidine, and their viability tested with trypan blue exclusion. MAPC were co-cultured with phenytoin and celecoxib at relevant clinical concentrations for 1 h and 24 h. Anti-inflammatory potency was tested using a stimulated rat splenocyte co-culture and ELISA for TNF-α production. MAPC were cultured under different osmolar concentrations and stained with propidium iodide for viability. Anti-inflammatory potency was tested by co-culture of MAPC with naïve lymphocytes activated by CD3/CD28 beads, and Click-iT® Plus EdU was used to quantify proliferation by flow cytometry. Results: The mean viability of the MAPC infused via needles was 95 ± 1%; no difference was seen with varying flow rate, but viability was notably reduced by chlorhexidine. MAPC function was not impaired by co-culture with phenytoin, celecoxib, or combination with both. Co-culture with phenytoin showed a decrease in TNF-α production as compared to the MAPC control. MAPC cultured at varying osmolar concentrations all had viabilities greater than 90% with no statistical difference between them. Co-culture of MAPC with CD3/CD28 activated PBMCs showed a significant reduction in proliferation as measured by EdU uptake. Discussion: Needle diameter, phenytoin, celecoxib, and a relevant range of osmolarities do not impair MAPC viability or anti-inflammatory potency in vitro.
