Thermoresponsive Substrates Used for the Growth and Controlled Differentiation of Human Mesenchymal Stem Cells

Abstract

This communication outlines the advances made in the development of thermoresponsive substrates for human mesenchymal stem cell (hMSC) expansion and subsequent controlled specific and multilineage differentiation from a previous study performed by this group. Previously, the development of an inexpensive and technically accessible method for hMSC expansion and harvesting was reported, using the solvent casting deposition method and thermoresponsive poly(N-isopropylacrylamide). Here, the logical continuation of this work is reported with the multipassage expansion of hMSCs with phenotypic maintenance followed by induced adipogenic, osteogenic, and chondrogenic differentiation. Interestingly, 1 μm thick solvent cast films are not only capable of hosting an expanding population of phenotypically preserved hMSCs similar to tissue culture plastic controls, but also the cells detached via temperature control better maintain their ability to differentiate compared to conventionally trypsinized cells. This approach to hMSC expansion and differentiation can be highly attractive to stem cell researchers where clinical therapies have seen a collective deviation away from the employment of animal derived products such as proteolytic trypsin.

Thermoresponsive and Active Functional Fiber Mats for Cultured Cell Recovery

“…Impressively, the cell proliferation ratio remained as high (88% relative to TCPS) as its original value found in the first round of the test suggesting that the GRGDS-CL fiber mat can be repeatedly used. This satisfactory feature has also been previously highlighted on solvent-cast PNIPAM film used as thermoresponsive platform for recovery of human mesenchymal stem cells …”

“…This satisfactory feature has also been previously highlighted on solvent-cast PNIPAM film used as thermoresponsive platform for recovery of human mesenchymal stem cells. 54 The ability of the GRGDS-CL fiber mat to yield detached cells with cell-to-cell junctions maintained can be observed by optical microscope. A representative brightfield image of a rolling cell sheet is shown in Figure S3, Supporting Information.…”

Surface immobilization of thermo-responsive poly(N-isopropylacrylamide) by simple entrapment in a 3-aminopropyltriethoxysilane network

“…They become hydrophilic when the temperature is below 32°C, and the polymer chains hydrate, promoting cell detachment. The use of pNIPAAm to harvest cells offers an advantage over using trypsin, which is known to cause some damage to cell membrane proteins and is reported to affect cell metabolism and differentiation[1–3]…”

Fabrication and Application of Photocrosslinked, Nanometer-Scale, Physically Adsorbed Films for Tissue Culture Regeneration

“…The detached cell sheets can then be transplanted for further cultivation in vitro or used for in vivo applications. A wide range of cell types have been successfully grown on pNIPAm based thermoresponsive surfaces to date and the number of studies dedicated to the refinement and improvement of this approach to cell sheet detachment is a testament to the attractiveness of the outcome . In spite of this, the widespread adoption of thermoresponsive surfaces for the production of cell sheets with a view to practical, clinical applications has been comparatively low.…”

“…A wide range of cell types have been successfully grown on pNIPAm based thermoresponsive surfaces to date and the number of studies dedicated to the refinement and improvement of this approach to cell sheet detachment is a testament to the attractiveness of the outcome. [3,[5][6][7][8] In spite of this, the widespread adoption of thermoresponsive surfaces for the production of cell sheets with a view to practical, clinical applications has been comparatively low. We, the authors, believe that this can be attributed to the fact that the preparation of such surfaces can be highly technologically complex and can require significant investment in terms of equipment and skills.…”