Abstract
Protein hydrogels represent ideal materials for advanced cell culture applications, including 3D-cultivation of even fastidious cells. Key properties of fully functional and, at the same time, economically successful cell culture materials are excellent biocompatibility and advanced fabrication processes allowing their easy production even on a large scale based on affordable compounds. Chemical crosslinking of bovine serum albumin (BSA) with N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC) in a water-in-oil emulsion with isoparaffinic oil as the continuous phase and sorbitan monooleate as surfactant generates micro-meter-scale spherical particles. They allow a significant simplification of an indispensable and laborious step in traditional cell culture workflows. This cell passaging (or splitting) to fresh culture vessels/flasks conventionally requires harsh trypsinization, which can be omitted by using the “trans-ferry-beads” presented here. When added to different pre-cultivated adherent cell lines, the beads are efficiently boarded by cells as passengers and can be easily transferred afterward for the embarkment of novel flasks. After this procedure, cells are perfectly viable and show normal growth behavior. Thus, the trans-ferry-beads not only may become extremely affordable as a final product but also may generally replace trypsinization in conventional cell culture, thereby opening new routes for the establishment of optimized and resource-efficient workflows in biological and medical cell culture laboratories.
Highlights
Hydrogels fabricated from biological sources such as proteins often termed “biogels”, have emerged as a versatile class of biocompatible and biodegradable materials for various applications such as cell culture technologies [10,11], smart wound dressings [13], and drug delivery [21]
Protein hydrogels based on blood proteins such as bovine or human serum albumin (BSA or HSA) even allow 3D cell culture applications, and they are comprehensively compatible with most cell lines
Crosslinking occurred by activating carboxyl groups of the bovine serum albumin (BSA) molecule with ethylcarbodiimide hydrochloride (EDC), forming an Oacylisourea active ester intermediate, which subsequently reacted with the amino groups of another or the same BSA molecule
Summary
Protein hydrogels were developed based on blood-derived proteins as building blocks, with serum albumins either from human or bovine origin being prominent examples. These proteins have crucial advantages for (biotechnological) applications, especially in the context of the field of cell culture/manipulation as they are considered per se biocompatible, in principle nonimmunogenic, and have proved their (enzymatic) biodegradability [10,11,12]. An additional advantage is that these proteins are economically extremely affordable, with bovine serum albumin (BSA) being even far less expensive than its human counterpart (HSA). Hydrogels have been developed based on these proteins, involving very simple crosslinking reactions with very inexpensive chemical crosslinkers such as four-armed amino reactive tetrakis(hydroxymethyl)phosphonium chloride (THPC) to form biocompatible and biodegradable hydrogel matrices [10,11,12,13,14,15,16]
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