Etd

Effect of Shear Stress on Cell Health and Secondary Metabolite Production in Taxus Plant Cell Culture

Public

Downloadable Content

open in viewer

Cancer is the second most common cause of death in many developed countries, making development of cancer treatments highly important. Paclitaxel is an FDA approved drug used for treatment of ovarian, breast, and lung cancer as well as Kaposi’s sarcoma. Due to high demand for paclitaxel, it is important to have a robust and scalable production method. Plant cell culture (PCC) of Taxus species is the current state-of-the-art production of paclitaxel on an industrial scale. One major drawback of producing paclitaxel using PCC is low and unstable yields. Thus, exploration of methods for increasing yield of paclitaxel in Taxus PCC is an ongoing field of research. When plant cells divide, they often form masses called aggregates that remain connected through their cell walls. Previous research in the Roberts laboratory has shown that large cellular aggregates produce less paclitaxel than smaller aggregates. We sought to investigate mechanical shearing of aggregates as a method for decreasing average aggregate size, which could be a method of increasing paclitaxel production. The three aims of this project are the following: determine how shearing affects Taxus secondary metabolism, including taxane, flavonoid, and phenolic biosynthesis, determine if stressors can make non-producing cell lines produce paclitaxel and design and perform tests with a novel device to implement continual shear stress on Taxus PCCs. Both long-term and continuous shear increased the levels of paclitaxel production, with methyl jasmonate elicited sheared 48.82A.3s (paclitaxel producing) cell cultures accumulating the highest levels overall. Cell culture growth was not impacted by shearing in both paclitaxel producing (48.82A.3s) and non-paclitaxel producing (P093XC) cell lines for both long-term and continuous shearing, which was consistent with that observed in other Taxus cell lines in previous studies. Finally, global secondary metabolism (total flavonoids and phenolics content) was not impacted by long-term shear for 48.82A.3s and P093XC cell cultures.

Creator
Contributors
Degree
Publisher
Identifier
  • etd-66101
Advisor
Defense date
Year
  • 2022
Date created
  • 2022-04-28
Resource type
Rights statement

Relations

In Collection:

Items

Items

Permanent link to this page: https://digital.wpi.edu/show/sj1395132