Therapeutic cloning applications for organ transplantation

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Abstract

A severe shortage of donor organs available for transplantation in the United States leaves patients suffering from diseased and injured organs with few treatment options. Scientists in the field of tissue engineering apply the principles of cell transplantation, material science, and engineering to construct biological substitutes that will restore and maintain normal function in diseased and injured tissues. Therapeutic cloning, where the nucleus from a donor cell is transferred into an enucleated oocyte in order to extract pluripotent embryonic stem cells, offers a potentially limitless source of cells for tissue engineering applications. The present chapter reviews recent advances that have occurred in therapeutic cloning and tissue engineering and describes applications of these new technologies that may offer novel therapies for patients with end-stage organ failure.

Introduction

The fields of regenerative medicine and tissue engineering aim to restore the form and function of damaged tissue and organs that have suffered from disease and injury. Many disorders, such as congenital anomalies, cancer, trauma, infection, inflammation, iatrogenic injuries, and other conditions, can lead to organ damage or loss and to the eventual need for reconstruction. It has been estimated that in the United States, one person in five reaching 65 years of age will receive temporary or permanent organ-replacement therapy during his or her remaining life span [1]. Furthermore, using the kidney as an example, over two million patients are projected to suffer from end-stage renal disease by 2010 and the aggregate health care costs for treating these patients has been estimated to be over $1 trillion dollars [2].

The majority of current reconstructive techniques rely on donor tissue for replacement; however, a shortage of donor tissue may limit these types of reconstruction, and usually significant morbidity is associated with the harvest procedure. Furthermore, the functional aspects of the damaged organ are rarely replaced by these reconstructive procedures, and they may even lead to complications because of the inherently different functional parameters of reconstructed tissue. Other potential sources of tissue include homologous tissues from cadavers, heterologous tissues from animal sources (bovine), and artificial materials (silicone, polyurethane, Teflon). Artificial devices made from these alternative sources were noted to be biocompatible and could provide structural replacement; however, the functional component of the original tissue was usually not recovered. While caution and disease prevention may never completely eradicate the incidence of disease and injury, regenerative medicine physicians and researchers hope to relieve the suffering of their patients from these unfortunate entities, and if possible, to repair or restore the damaged tissue with the hope of regenerating essentially normal body parts.

Organ transplantation was one of the first methods to restore form and function in modern medicine. In 1955, Murray performed the first successful organ transplant (kidney), and in the early 1960s, he performed the first allogeneic kidney transplantation from a genetically dissimilar donor into an unrelated recipient [3]. As one of the first procedures to overcome the immunologic barrier, this revolutionary procedure marked the modern era in which transplantation could be used as means of therapy for diseased and injured organs. Since then, advances in immunosuppressive medications, in the matching of similar donors to recipients, and in the treatment of rejections have resulted in thousands of patients each year undergoing successful transplantation of donor organs. In 2001, over 23 000 patients received a transplanted organ in the United States [4].

However, despite the advances in transplantation over the past 50 years, a severe shortage of donor organs limits the availability of this treatment, such that in 2001, nearly 80 000 patients were awaiting a donor organ, and over 6000 patients were reported to have died while awaiting an organ transplant [4]. This has spawned the search for alternate therapies, such as therapeutic cloning, that can be used in regenerative medicine applications.

Regenerative medicine has the potential to benefit from cloning technology. Cloning techniques are already widely utilized in the modern world, where gardeners perform plant cuttings and commercial companies propagate desirable plant strains and animal breeds [5]. However, since the birth of Dolly as the first animal cloned from an adult somatic cell in 1997, tremendous further interest in cloning has arisen, and cloning has been envisioned as having utility in medical applications as well.

Section snippets

Definition of therapeutic cloning

Nuclear cloning, which has also been called nuclear transplantation and nuclear transfer, involves the introduction of a nucleus from a donor cell into an enucleated oocyte to generate an embryo with a genetic makeup identical to that of the donor.

While there has been tremendous interest in the field of nuclear cloning since the birth of Dolly in 1997, the first successful nuclear transfer was reported over 50 years ago by Briggs and King [6]. Cloned frogs, which were the first vertebrates

Stem cells and therapeutic cloning

Most current strategies for tissue/organ replacement therapy depend upon a sample of autologous cells from the diseased organ of the host. However, for many patients with extensive end-stage organ failure, a tissue biopsy may not yield enough normal cells for expansion and eventual transplantation. In these situations, embryonic stem cells are envisioned as an alternative source of cells from which the desired tissue can be derived. Combining the techniques learned in tissue engineering over

Further improvements in somatic cell nuclear transfer technology

While promising, somatic cell nuclear transfer technology has certain limitations that require further improvements before therapeutic cloning can be applied widely in replacement therapy.

Currently, the efficiency of the overall cloning process is low. The majority of embryos derived from animal cloning do not survive after implantation [38], [39], [40]. In practical terms, multiple nuclear transfers must be performed in order to produce one live offspring for animal cloning applications. The

Tissue engineering principles

After an adequate amount of cells are expanded from a cloned source, tissue engineering will be required to produce transplantable tissue or organs. The field of tissue engineering has emerged over the past 40 years to address the shortcomings of previous replacement therapies. Scientists in this relatively new field aim to combine the principles of cell transplantation, material science, and engineering to construct biological substitutes that will restore and maintain normal function in

Applications of therapeutic cloning

We applied the principles of both tissue engineering and therapeutic cloning in an effort to produce genetically identical renal and cardiac muscle tissue in a large animal model, the cow (Bos taurus) [78]. Unfertilized bovine donor oocytes were retrieved from abbatoir-derived ovaries, and the nuclear material from these eggs was removed by mechanical enucleation 18–22 h after maturation. Complete enucleation of the metaphase plate was confirmed with bisBenzimide dye under fluorescence

References (92)

  • A. Atala et al.

    Implantation in vivo and retrieval of artificial structures consisting of rabbit and human urothelium and human bladder muscle

    J Urol

    (1993)
  • A. Atala et al.

    A novel inert collagen matrix for hypospadias repair

    J Urol

    (1999)
  • A. Atala et al.

    Endoscopic treatment of vesicoureteral reflux with a chondrocyte-alginate suspension

    J Urol

    (1994)
  • B.G. Cilento et al.

    Phenotypic and cytogenetic characterization of human bladder urothelia expanded in vitro

    J Urol

    (1994)
  • D.O. Fauza et al.

    Videofetoscopically assisted fetal tissue engineering: bladder augmentation

    J Pediatr Surg

    (1998)
  • D.O. Fauza et al.

    Videofetoscopically assisted fetal tissue engineering: skin replacement

    J Pediatr Surg

    (1998)
  • R.T. Kershen et al.

    New advances in injectable therapies for the treatment of incontinence and vesicoureteral reflux

    Urol Clin North Am

    (1999)
  • H.J. Park et al.

    Reconstitution of human corporal smooth muscle and endothelial cells in vivo

    J Urol

    (1999)
  • J.J. Yoo et al.

    A novel gene delivery system using urothelial tissue engineered neo-organs

    J Urol

    (1997)
  • J.J. Yoo et al.

    Cartilage rods as a potential material for penile reconstruction

    J Urol

    (1998)
  • M. Liebert et al.

    Expression of mal is associated with urothelial differentiation in vitro: identification by differential display reverse-transcriptase polymerase chain reaction

    Differentiation

    (1997)
  • S.D. Scriven et al.

    Reconstitution of human urothelium from monolayer cultures

    J Urol

    (1997)
  • B.S. Kim et al.

    Development of biocompatible synthetic extracellular matrices for tissue engineering

    Trends Biotechnol

    (1998)
  • J.E. Bergsma et al.

    In vivo degradation and biocompatibility study of in vitro pre-degraded as-polymerized polyactide particles

    Biomaterials

    (1995)
  • R.O. Hynes

    Integrins: versatility, modulation, and signaling in cell adhesion

    Cell

    (1992)
  • M.J. Lysaght et al.

    Demographic scope and economic magnitude of contemporary organ replacement therapies

    ASAIO J

    (2000)
  • M.J. Lysaght

    Maintenance dialysis population dynamics: current trends and long-term implications

    J Am Soc Nephrol

    (2002)
  • J.E. Murray et al.

    Renal homotransplantation in identical twins

    Surg Forum

    (1955)
  • UNOS. 2002 OPTN/SRTR Annual Report. www.optn.org...
  • J.B. Gurdon et al.

    The future of cloning

    Nature

    (1999)
  • R. Briggs et al.

    Transplantation of living nuclei from blastula cells into enucleated frogs’ eggs

    Proc Natl Acad Sci USA

    (1952)
  • K.H. Campbell et al.

    Sheep cloned by nuclear transfer from a cultured cell line

    Nature

    (1996)
  • I. Wilmut et al.

    Viable offspring derived from fetal and adult mammalian cells

    Nature

    (1997)
  • J.B. Cibelli et al.

    Cloned transgenic calves produced from non-quiescent fetal fibroblasts

    Science

    (1998)
  • A. Baguisi et al.

    Production of goats by somatic cell nuclear transfer

    Nat Biotechnol

    (1999)
  • C.L. Keefer et al.

    Production of cloned goats after nuclear transfer using adult somatic cells

    Biol Reprod

    (2002)
  • T. Wakayama et al.

    Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei

    Nature

    (1998)
  • J. Betthauser et al.

    Production of cloned pigs from in vitro systems

    Nat Biotechnol

    (2000)
  • I.A. Polejaeva et al.

    Cloned pigs produced by nuclear transfer from adult somatic cells

    Nature

    (2000)
  • A. Onishi et al.

    Pig cloning by microinjection of fetal fibroblast nuclei

    Science

    (2000)
  • P.A. De Sousa et al.

    Somatic cell nuclear transfer in the pig: control of pronuclear formation and integration with improved methods for activation and maintenance of pregnancy

    Biol Reprod

    (2002)
  • B. Vogelstein et al.

    Genetics. Please don't call it cloning!

    Science

    (2002)
  • K. Hochedlinger et al.

    Nuclear transplantation, embryonic stem cells, and the potential for cell therapy

    New Engl J Med

    (2003)
  • R.P. Lanza et al.

    The ethical reasons for stem cell research

    Science

    (2001)
  • R.P. Lanza et al.

    Prospects for the use of nuclear transfer in human transplantation

    Nat Biotechnol

    (1999)
  • A.H. Brivanlou et al.

    Stem cells. Setting standards for human embryonic stem cells

    Science

    (2003)
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