Tissue Engineering Using Human Embryonic Stem Cells

Stem Cell Center, Bruce Rappaport Faculty of Medicine, Technion - Israel, Haifa, Israel.
Methods in Enzymology (Impact Factor: 2.09). 02/2006; 420:303-15. DOI: 10.1016/S0076-6879(06)20014-4
Source: PubMed


The possibility of using stem cells for tissue engineering has greatly encouraged scientists to design new platforms in the field of regenerative and reconstructive medicine. Stem cells have the ability to rejuvenate and repair damaged tissues and can be derived from both embryonic and adult sources. Among cell types suggested as a cell source for tissue engineering (TE), human embryonic stem cells (hESCs) are one of the most promising candidates. Isolated from the inner cell mass of preimplantation stage blastocysts, they possess the ability to differentiate into practically all adult cell types. In addition, their unlimited self-renewal capacity enables the generation of sufficient amount of cells for cell-based TE applications. Yet, several important challenges are to be addressed, such as the isolation of the desired cell type and gaining control over its differentiation and proliferation. Ultimately, combing scaffolding and bioactive stimuli, newly designed bioengineered constructs, could be assembled and applied to various clinical applications. Here we define the culture conditions for the derivation of connective tissue lineage progenitors, design strategies, and highlight the special considerations when using hESCs for TE applications.

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    • "Due to their potentially unlimited capacity for self-renewal and unique developmental potential to differentiate into all somatic cell types of the human body, hES cells have opened a new door for drug discovery, regenerative medicine, and tissue replacement after injury or disease.1, 2 However, an essential prerequisite for successful applications of hES cells is to develop efficient cryopreservation methods to overcome the current low cell recovery rate after cryopreservation. "
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    ABSTRACT: Due to widespread applications of human embryonic stem (hES) cells, it is essential to establish effective protocols for cryopreservation and subsequent culture of hES cells to improve cell recovery. We have developed a new protocol for cryopreservation of dissociated hES cells and subsequent culture. We examined the effects of new formula of freezing solution containing 7.5% dimethylsulfoxide (DMSO) (v/v %) and 2.5% polyethylene glycol (PEG) (w/v %) on cell survival and recovery of hES cells after cryopreservation, and further investigated the role of the combination of Rho-associated kinase (ROCK) inhibitor and p53 inhibitor on cell recovery during the subsequent culture. Compared with the conventional slow-freezing method which uses 10% DMSO as a freezing solution and then cultured in the presence of ROCK inhibitor at the first day of culture, we found out that hES cell recovery was significantly enhanced by around 30 % (P < 0.05) by the new freezing solution. Moreover, at the first day of post-thaw culture, the presence of 10 microM ROCK inhibitor (Y-27632) and 1 microM pifithrin-mu together further significantly improved cell recovery by around 20% (P < 0.05) either for feeder-dependent or feeder-independent culture. hES cells remained their undifferentiated status after using this novel protocol for cryopreservation and subsequent culture. Furthermore, this protocol is a scalable cryopreservation method for handling large quantities of hES cells.
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    • "Arguments can be made for a stem cell source being optimal for applications in regenerative therapies, and, importantly, studies are needed to define the necessity of stem cells in such endeavors. There are more data available regarding cartilage tissue engineering, and those data support the need for the presence of cells (chondrocytes or stem cells) in a graft composite , but similar data are less abundant for tendon regenerative studies [17] [18] [19]. "
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    ABSTRACT: After tendon injury, the scar tissue that replaces the damaged tendon results in a substantial risk for reinjury. The goal of regenerative therapies is to restore normal structural architecture and biomechanical function to an injured tissue. Successful restoration processes for any tissue are thought to recapitulate those of development, in which there are spatial and temporal interactions between scaffold, growth factors, and cell populations.
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