Methods for isolating, expanding, and characterizing umbilical cord mesenchymal stromal cells and their in vitro metabolism

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dc.contributor.author Smith, Joseph Robert
dc.date.accessioned 2017-04-20T21:38:46Z
dc.date.available 2017-04-20T21:38:46Z
dc.date.issued 2017-05-01 en_US
dc.identifier.uri http://hdl.handle.net/2097/35462
dc.description.abstract Mesenchymal stromal cells (MSCs) derived from the umbilical cord (UC-MSCs) have therapeutic applications and are studied to understand their potential uses and immunomodulatory properties. Research must identify good manufacturing process (GMP) compliant methods to isolate and expand UC-MSCs. In addition, MSCs metabolism characteristics in culture are unknown, warranting further investigation. Viability of MSCs decreases after cryopreservation, which is detrimental to clinical translation. Previously published methods used to isolate MSCs from the umbilical cord included open dissection steps and xenogeneic components. Here, I developed improved methods by eliminating dissection which reduces contamination risks. Instead, I used the whole umbilical cord and Miltenyi dissociator tubes to mechanically and enzymatically dissociate cells in a closed system. Xenogeneic components were decreased by using medium containing pooled human platelet lysate instead of fetal bovine serum. The cell numbers isolated from umbilical cord averaged 2.68 x 10⁵ per cm, which represents greater than 20 fold improvement over the previous method. Moreover, expansion cell numbers were increased using 10% pooled human platelet lysate supplemented media. The UC-MSCs generated here met the International Society of Cell Therapy (ISCT) definition of MSCs. Metabolism characteristics of MSCs indicated that glucose was the critical metabolite, maintaining cells longer in culture than glutamine. Cell death followed depletion of glucose, too. Finally, the average viability after thawing cryopreserved MSCs was more than 95%, higher than previous methods. The improvements I introduced to our methodology could speed clinical translation of MSCs as an allogeneic cellular therapy en_US
dc.language.iso en_US en_US
dc.publisher Kansas State University en
dc.subject Mesenchymal stromal cells en_US
dc.subject Stem cells en_US
dc.subject Umbilical cord en_US
dc.subject Platelet lysate en_US
dc.subject Isolation methods en_US
dc.subject Cryopreservation en_US
dc.title Methods for isolating, expanding, and characterizing umbilical cord mesenchymal stromal cells and their in vitro metabolism en_US
dc.type Thesis en_US
dc.description.degree Master of Science in Biomedical Sciences en_US
dc.description.level Masters en_US
dc.description.department Department of Anatomy and Physiology en_US
dc.description.advisor Mark L. Weiss en_US
dc.date.published 2017 en_US
dc.date.graduationmonth May en_US


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