Cell culture platform with mechanical conditioning and  nondamaging cellular detachment.
Cells implanted after injury may remodel undesirably with improper mechanical stimulation from surrounding tissue. Proper conditioning of tissue engineered constructs before implantation can lead to suitable tissue architectures, along with an extracellular matrix (ECM) environment that more closely mimics native tissue. Additionally, cell implantation without bulky polymeric scaffolding is often desirable. Previous researchers have created devices capable of applying mechanical forces to cells (e.g., stretch) but cellular removal from these devices, such as by trypsin, often results in irreversible damage. Conversely, devices are available that can detach intact cells, but these are inelastic, nonstretchable substrates. We have created a cell culture platform that allows for mechanical conditioning and then subsequent nondamaging detachment of those cells. We have modified silicone culture surfaces, to incorporate thermally responsive polymers of N-isopropylacrylamide (NIPAAm) to create an elastic substrate that can also change surface properties with temperature change. A copolymer of NIPAAm and 10percent w/w acrylic acid (AAc) was conjugated to an amine-bonded silicone surface through carbodiimide chemistry. Cells were able to attach to the resulting surfaces at 37 degreeC and showed detachment by rounded morphology at 25degreeC. Following mechanical stretching, cells were still able to spontaneously detach from these modified silicone surfaces with temperature change.
Lee
Elaine L
EL
Department of Biomedical Engineering, Case Western Reserve University, Wickenden 220, 10900 Euclid Avenue,Cleveland, Ohio 44106, USA.
von Recum
Horst A
HA
D000178:Acrylamides / Q000737:chemistry; D000179:Acrylates / Q000737:chemistry; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry / Q000378:metabolism; D019149:Bioreactors; D002448:Cell Adhesion / Q000502:physiology*; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism; D008422:Materials Testing; D051379:Mice; D015394:Molecular Structure; D041681:NIH 3T3 Cells; D011108:Polymers / Q000737:chemistry; D012828:Silicones / Q000737:chemistry; D013314:Stress, Mechanical; D013499:Surface Properties; D013696:Temperature; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods
D000178:Acrylamides; D000179:Acrylates; D001672:Biocompatible Materials; D011108:Polymers; D012828:Silicones; C067295:N-isopropylacrylamide; C036658:acrylic acid