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Analysis of host-mediated repair mechanisms after human CNS-stem cell transplantation for spinal cord injury: correlation of engraftment with recovery.

Hooshmand MJ, Sontag CJ, Uchida N, Tamaki S, Anderson AJ, Cummings BJ - PLoS ONE (2009)

Bottom Line: Stereological quantification of human cells using a human-specific cytoplasmic marker demonstrated successful cell engraftment, survival, migration and limited proliferation in all hCNS-SCns transplanted animals.Biochemical analyses supplemented stereological data supporting the absence of neural stem-cell mediated host repair.However, linear regression analysis of the number of engrafted hCNS-SCns vs. the number of errors on a horizontal ladder beam task revealed a strong correlation between these variables (r = -0.78, p<0.05), suggesting that survival and engraftment were directly related to a quantitative measure of recovery.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy and Neurobiology, University of California Irvine, Irvine, CA, USA.

ABSTRACT

Background: Human central nervous system-stem cells grown as neurospheres (hCNS-SCns) self-renew, are multipotent, and have potential therapeutic applications following trauma to the spinal cord. We have previously shown locomotor recovery in immunodeficient mice that received a moderate contusion spinal cord injury (SCI) and hCNS-SCns transplantation 9 days post-injury (dpi). Engrafted hCNS-SCns exhibited terminal differentiation to myelinating oligodendrocytes and synapse-forming neurons. Further, selective ablation of human cells using Diphtheria toxin (DT) abolished locomotor recovery in this paradigm, suggesting integration of human cells within the mouse host as a possible mechanism for the locomotor improvement. However, the hypothesis that hCNS-SCns could alter the host microenvironment as an additional or alternative mechanism of recovery remained unexplored; we tested that hypothesis in the present study.

Methods and findings: Stereological quantification of human cells using a human-specific cytoplasmic marker demonstrated successful cell engraftment, survival, migration and limited proliferation in all hCNS-SCns transplanted animals. DT administration at 16 weeks post-transplant ablated 80.5% of hCNS-SCns. Stereological quantification for lesion volume, tissue sparing, descending serotonergic host fiber sprouting, chondroitin sulfate proteoglycan deposition, glial scarring, and angiogenesis demonstrated no evidence of host modification within the mouse spinal cord as a result of hCNS-SCns transplantation. Biochemical analyses supplemented stereological data supporting the absence of neural stem-cell mediated host repair. However, linear regression analysis of the number of engrafted hCNS-SCns vs. the number of errors on a horizontal ladder beam task revealed a strong correlation between these variables (r = -0.78, p<0.05), suggesting that survival and engraftment were directly related to a quantitative measure of recovery.

Conclusions: Altogether, the data suggest that the locomotor improvements associated with hCNS-SCns transplantation were not due to modifications within the host microenvironment, supporting the hypothesis that human cell integration within the host circuitry mediates functional recovery following a 9 day delayed transplant.

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hCNS-SCns engraftment directly correlates with a quantitative measure of behavior, but not other measures of histological recovery.A: Linear regression analysis revealed a significant negative correlation between hCNS-SCns engraftment and the number of errors made on the horizontal ladderbeam task (Pearson r = −0.78, p = 0.038, 2-tailed t-test). B-D: Linear regression analyses for the estimated number of hCNS-SCns and other measures of host recovery revealed no significant correlations between cell engraftment and lesion volume (B) (Pearson r = −0.70, p = 0.08, 2-tailed t-test), volume of spared tissue (B) (Pearson r = 0.42, p = 0.34, 2-tailed t-test), serotonergic fiber sprouting (C) (Pearson r = 0.16, p = 0.77, 2-tailed t-test), NG2 area (D) (Pearson r = 0.16, p = 0.77, 2-tailed t-test), and area of the GFAP astroglial scar (D) (Pearson r = −0.69, p = 0.08, 2-tailed t-test). * denotes p<0.05.
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pone-0005871-g011: hCNS-SCns engraftment directly correlates with a quantitative measure of behavior, but not other measures of histological recovery.A: Linear regression analysis revealed a significant negative correlation between hCNS-SCns engraftment and the number of errors made on the horizontal ladderbeam task (Pearson r = −0.78, p = 0.038, 2-tailed t-test). B-D: Linear regression analyses for the estimated number of hCNS-SCns and other measures of host recovery revealed no significant correlations between cell engraftment and lesion volume (B) (Pearson r = −0.70, p = 0.08, 2-tailed t-test), volume of spared tissue (B) (Pearson r = 0.42, p = 0.34, 2-tailed t-test), serotonergic fiber sprouting (C) (Pearson r = 0.16, p = 0.77, 2-tailed t-test), NG2 area (D) (Pearson r = 0.16, p = 0.77, 2-tailed t-test), and area of the GFAP astroglial scar (D) (Pearson r = −0.69, p = 0.08, 2-tailed t-test). * denotes p<0.05.

Mentions: Linear regression analysis revealed a significant negative correlation (Pearson r: r = −0.78, p = 0.038, 2-tailed t-test) between the estimated number of engrafted hCNS-SCns and the number of errors made on the horizontal ladderbeam (Fig. 11A). Interestingly, a positive but non-significant, correlation (Pearson r: r = 0.49, p = 0.26, 2-tailed t-test) was found between the estimated number of hFb and the number of errors made on the ladderbeam task (Fig. 12A), suggesting a trend for decreased behavioral recovery with an increase in hFb engraftment. These data extend and confirm our previous findings and suggest that hCNS-SCns, but not hFb engraftment is critical for locomotor recovery.


Analysis of host-mediated repair mechanisms after human CNS-stem cell transplantation for spinal cord injury: correlation of engraftment with recovery.

Hooshmand MJ, Sontag CJ, Uchida N, Tamaki S, Anderson AJ, Cummings BJ - PLoS ONE (2009)

hCNS-SCns engraftment directly correlates with a quantitative measure of behavior, but not other measures of histological recovery.A: Linear regression analysis revealed a significant negative correlation between hCNS-SCns engraftment and the number of errors made on the horizontal ladderbeam task (Pearson r = −0.78, p = 0.038, 2-tailed t-test). B-D: Linear regression analyses for the estimated number of hCNS-SCns and other measures of host recovery revealed no significant correlations between cell engraftment and lesion volume (B) (Pearson r = −0.70, p = 0.08, 2-tailed t-test), volume of spared tissue (B) (Pearson r = 0.42, p = 0.34, 2-tailed t-test), serotonergic fiber sprouting (C) (Pearson r = 0.16, p = 0.77, 2-tailed t-test), NG2 area (D) (Pearson r = 0.16, p = 0.77, 2-tailed t-test), and area of the GFAP astroglial scar (D) (Pearson r = −0.69, p = 0.08, 2-tailed t-test). * denotes p<0.05.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2690693&req=5

pone-0005871-g011: hCNS-SCns engraftment directly correlates with a quantitative measure of behavior, but not other measures of histological recovery.A: Linear regression analysis revealed a significant negative correlation between hCNS-SCns engraftment and the number of errors made on the horizontal ladderbeam task (Pearson r = −0.78, p = 0.038, 2-tailed t-test). B-D: Linear regression analyses for the estimated number of hCNS-SCns and other measures of host recovery revealed no significant correlations between cell engraftment and lesion volume (B) (Pearson r = −0.70, p = 0.08, 2-tailed t-test), volume of spared tissue (B) (Pearson r = 0.42, p = 0.34, 2-tailed t-test), serotonergic fiber sprouting (C) (Pearson r = 0.16, p = 0.77, 2-tailed t-test), NG2 area (D) (Pearson r = 0.16, p = 0.77, 2-tailed t-test), and area of the GFAP astroglial scar (D) (Pearson r = −0.69, p = 0.08, 2-tailed t-test). * denotes p<0.05.
Mentions: Linear regression analysis revealed a significant negative correlation (Pearson r: r = −0.78, p = 0.038, 2-tailed t-test) between the estimated number of engrafted hCNS-SCns and the number of errors made on the horizontal ladderbeam (Fig. 11A). Interestingly, a positive but non-significant, correlation (Pearson r: r = 0.49, p = 0.26, 2-tailed t-test) was found between the estimated number of hFb and the number of errors made on the ladderbeam task (Fig. 12A), suggesting a trend for decreased behavioral recovery with an increase in hFb engraftment. These data extend and confirm our previous findings and suggest that hCNS-SCns, but not hFb engraftment is critical for locomotor recovery.

Bottom Line: Stereological quantification of human cells using a human-specific cytoplasmic marker demonstrated successful cell engraftment, survival, migration and limited proliferation in all hCNS-SCns transplanted animals.Biochemical analyses supplemented stereological data supporting the absence of neural stem-cell mediated host repair.However, linear regression analysis of the number of engrafted hCNS-SCns vs. the number of errors on a horizontal ladder beam task revealed a strong correlation between these variables (r = -0.78, p<0.05), suggesting that survival and engraftment were directly related to a quantitative measure of recovery.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy and Neurobiology, University of California Irvine, Irvine, CA, USA.

ABSTRACT

Background: Human central nervous system-stem cells grown as neurospheres (hCNS-SCns) self-renew, are multipotent, and have potential therapeutic applications following trauma to the spinal cord. We have previously shown locomotor recovery in immunodeficient mice that received a moderate contusion spinal cord injury (SCI) and hCNS-SCns transplantation 9 days post-injury (dpi). Engrafted hCNS-SCns exhibited terminal differentiation to myelinating oligodendrocytes and synapse-forming neurons. Further, selective ablation of human cells using Diphtheria toxin (DT) abolished locomotor recovery in this paradigm, suggesting integration of human cells within the mouse host as a possible mechanism for the locomotor improvement. However, the hypothesis that hCNS-SCns could alter the host microenvironment as an additional or alternative mechanism of recovery remained unexplored; we tested that hypothesis in the present study.

Methods and findings: Stereological quantification of human cells using a human-specific cytoplasmic marker demonstrated successful cell engraftment, survival, migration and limited proliferation in all hCNS-SCns transplanted animals. DT administration at 16 weeks post-transplant ablated 80.5% of hCNS-SCns. Stereological quantification for lesion volume, tissue sparing, descending serotonergic host fiber sprouting, chondroitin sulfate proteoglycan deposition, glial scarring, and angiogenesis demonstrated no evidence of host modification within the mouse spinal cord as a result of hCNS-SCns transplantation. Biochemical analyses supplemented stereological data supporting the absence of neural stem-cell mediated host repair. However, linear regression analysis of the number of engrafted hCNS-SCns vs. the number of errors on a horizontal ladder beam task revealed a strong correlation between these variables (r = -0.78, p<0.05), suggesting that survival and engraftment were directly related to a quantitative measure of recovery.

Conclusions: Altogether, the data suggest that the locomotor improvements associated with hCNS-SCns transplantation were not due to modifications within the host microenvironment, supporting the hypothesis that human cell integration within the host circuitry mediates functional recovery following a 9 day delayed transplant.

Show MeSH
Related in: MedlinePlus