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Preparation of thermosensitive PNIPAm‐based copolymer coated cytodex 3 microcarriers for efficient nonenzymatic cell harvesting during 3D culturing
Author(s) -
Darge Haile F.,
Chuang ShunHao,
Lai JuinYih,
Lin ShuianYin,
Tsai HsiehChih
Publication year - 2021
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.27885
Subject(s) - lower critical solution temperature , microcarrier , materials science , polymer , chemical engineering , copolymer , adhesion , scanning electron microscope , cell adhesion , polymer chemistry , chemistry , cell , composite material , biochemistry , engineering
Enzymatic detachment of cells might damage important features and functions of cells and could affect subsequent cell‐based applications. Therefore, nonenzymatic cell detachment using thermosensitive polymer matrix is necessary for maintaining cell quality after harvesting. In this study, we prepared thermosensitive PNIPAm‐ co ‐AAc‐ b ‐PS and PNIPAm‐ co ‐AAm‐ b ‐PS copolymers and low critical solution temperature (LCST) was tuned near to body temperature. Then, spin coated polymer films were prepared for cell adhesion and thermal‐induced cell detachment. The alpha‐step analysis and scanning electron microscope image of the films suggested that the thickness of the films depends on the molecular weight and concentration which ranged from 206 to 1330 nm for PNIPAm‐ co ‐AAc‐ b ‐PS and 97.5–497 nm for PNIPAm‐ co ‐AAm‐ b ‐PS. The contact angles of the films verified that the polymer surface was moderately hydrophilic at 37°C. Importantly, RAW264.7 cells were convincingly proliferated on the films to a confluent of >80% within 48 h and abled to detach by reducing the temperature. 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, PNIPAm‐ co ‐AAm‐ b ‐PS (5%w/v) was further used to coat commercial cytodex 3 microcarriers for 3D cell culturing and interestingly enhanced cell detachment with preserved potential of recovery was observed at a temperature of below LCST. Thus, surface modification of microcarriers with thermosensitive PNIPAm‐ co ‐AAm‐ b ‐PS could be vital strategy for nonenzymatic cell detachment and to achieve adequate number of cells with maximum cell viability and functionality.