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Preparation of thermosensitive PNIPAm-based copolymer coated cytodex 3 microcarriers for efficient non-enzymatic cell harvesting during 3D culturing
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  • Haile Darge,
  • Shun-Hao Chuang,
  • Juin-Yih Lai,
  • Shuian-Yin Lin,
  • Hsieh-Chih Tsai
Haile Darge
National Taiwan University of Science and Technology
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Shun-Hao Chuang
National Taiwan University of Science and Technology
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Juin-Yih Lai
National Taiwan University of Science and Technology
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Shuian-Yin Lin
Industrial Technology Research Institute
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Hsieh-Chih Tsai
National Taiwan University of Science and Technology
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Abstract

Enzymatic detachment of cells might damage important features of cells and could affect subsequent function of cells in various applications. Therefore, non-enzymatic cell detachment using thermosensitive polymer matrix is necessary for maintaining cell quality after harvesting. In this study, we synthesized thermosensitive PNIPAm-co-AAc-b-PS and PNIPAm-co-AAm-b-PS copolymers and LCST was tuned near to body temperature. Then, polymer solutions (5% w/v, 10% w/v, and 20% w/v) were spin coated to prepare films for cell adhesion and thermal-induced cell detachment. The apha-step analysis and SEM image of the films suggested that the thickness of the films depends on the molecular weight and concentration which ranged from 206 nm to 1330 nm for PNIPAm-co-AAc-b-PS and 97.5 nm to 497 nm for PNIPAm-co-AAm-b-PS. The contact angles of the films verified that the polymer surface was moderately hydrophilic at 37°C. From cell attachment and detachment studies, RAW264.7 cells, were convincingly proliferated on the films to a confluent of >80 % within 48 days. However, relatively more cells were grown on PNIPAm-co-AAm-b-PS (5%w/v) films and thermal-induced cell detachment was more abundant in this formulation. As a result, commercial cytodex 3 microcarrier was coated with PNIPAm-co-AAm-b-PS (5%w/v) and interestingly enhanced cell detachment with preserved potential of recovery was observed at low temperature during 3D culturing. Thus, surface modification of microcarriers with PNIPAm-co-AAm-b-PS could be vital strategy for non-enzymatic cell dissociation and able to achieve adequate number of cells with maximum cell viability, and functionality for various cell-based applications. Keywords: surface coated microcarriers; thermosensitive polymer; non-enzymatic cell detachment

Peer review status:UNDER REVIEW

10 Mar 2021Submitted to Biotechnology and Bioengineering
10 Mar 2021Submission Checks Completed
10 Mar 2021Assigned to Editor
24 Mar 2021Reviewer(s) Assigned
18 Apr 2021Review(s) Completed, Editorial Evaluation Pending
18 Apr 2021Editorial Decision: Revise Major
30 Apr 20211st Revision Received
30 Apr 2021Submission Checks Completed
30 Apr 2021Assigned to Editor
18 May 2021Reviewer(s) Assigned
07 Jun 2021Editorial Decision: Revise Minor
07 Jun 2021Review(s) Completed, Editorial Evaluation Pending
09 Jun 20212nd Revision Received
10 Jun 2021Submission Checks Completed
10 Jun 2021Assigned to Editor
10 Jun 2021Reviewer(s) Assigned