Development of a Model of Sacrocaudal Spinal Cord Injury in Cloned Yucatan MiniPigs for Cellular Transplantation Research

Abstract

Research into transplantation strategies to treat spinal cord injury (SCI) is frequently performed in rodents, but translation of results to clinical patients can be poor and a large mammalian model of severe SCI is needed. The pig has been considered an optimal model species in which to perform preclinical testing, and the Yucatan minipig can be cloned successfully utilizing somatic cell nuclear transfer (SCNT). However, induction of paralysis in pigs poses significant welfare and nursing challenges. The present study was conducted to determine whether Yucatan SCNT clones could be used to develop an SCI animal model for cellular transplantation research. First, we demonstrated that transection of the sacrocaudal spinal cord in Yucatan SCNT clones produces profound, quantifiable neurological deficits restricted to the tail. We then established that neurospheres could be isolated from brain tissue of green fluorescence protein (GFP) transfected SCNT clones. Finally, we confirmed survival of transplanted GFP-expressing neural stem cells in the SCI lesion and their differentiation into glial and neuronal lineages for up to 4 weeks without immunosuppression. We conclude that this model of sacrocaudal SCI in Yucatan SCNT clones represents a powerful research tool to investigate the effect of cellular transplantation on axonal regeneration and functional recovery.

Alternating Current Electric Fields of Varying Frequencies: Effects on Proliferation and Differentiation of Porcine Neural Progenitor Cells

“…Porcine neurospheres were generated using a protocol based on previously reported neurosphere culture methods (Lim et al, 2010). Briefly, after euthanasia, brains were removed and placed into a balanced salt solution (Gibco-BRL, Gaithersburg, MD, USA) and then grossly dissected, minced, and enzymatically digested to produce single cells.…”

Effect of kidney-reinforcing and marrow-beneficial Chinese medicine on bone metabolism-related factors following spinal cord injury in rats

“…The advantage of using this surgical procedure is that it operates at the correct anatomical position and is consistent with the degree of injury. The operation results in motor and sensory function loss below the cross section, causing dysfunction that is attributed to the primary injury of the spinal cord and diminishing the risk of human error (14).…”

Translational spinal cord injury research

“…Depending upon the specific questions being addressed, other large animal models, such as the cat (Tester and How-land, 2008; Cote et al, 2010) and mini-pig (Zurita et al, 2008; Lim et al, 2010; Kuluz et al, 2010; Navarro et al, 2012; Guest et al, 2011; Wewetzer et al, 2011), may serve as alternatives to nonhuman primates. Most notably, cat models of contusion and compression injuries have a long preclinical history related to neuroprotection and cell replacement therapies (Anderson et al, 1985; Hall and Braughler, 1986; Anderson et al, 1995) as well as an extensive background of spinal neurophysiology and neuroanatomical characterization.…”

“…Presently, there are no established standards for defining the best immunosuppression protocols with the fewest possible contraindications (Barker and Widner, 2004). Some reports suggest neural precursors are not highly immunogenic and do not require immunosuppression (Lim et al, 2010). In fact, a recently initiated trial (Pilot Investigation of Stem Cells in Stroke) in Glasgow (ReNeuron, Ltd.) is not employing any form of immunosuppression for that reason.…”