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Intranasal administration of human MSC for ischemic brain injury in the mouse: in vitro and in vivo neuroregenerative functions.

Donega V, Nijboer CH, Braccioli L, Slaper-Cortenbach I, Kavelaars A, van Bel F, Heijnen CJ - PLoS ONE (2014)

Bottom Line: Our results show that 2×10(6) hMSCs decrease lesion volume, improve motor behavior, and reduce scar formation and microglia activity.This is suggested by our finding that CXCL10 is significantly upregulated at 10 days following HI, but not at 17 days after HI, a time when MSCs no longer reach the lesion when given intranasally.The results described in this work also tempt us to contemplate hMSCs not only as a potential treatment option for neonatal encephalopathy, but also for a plethora of degenerative and traumatic injuries of the nervous system.

View Article: PubMed Central - PubMed

Affiliation: Lab. of Neuroimmunology and Developmental Origins of Disease, University Medical Center Utrecht, Utrecht, The Netherlands.

ABSTRACT
Intranasal treatment with C57BL/6 MSCs reduces lesion volume and improves motor and cognitive behavior in the neonatal hypoxic-ischemic (HI) mouse model. In this study, we investigated the potential of human MSCs (hMSCs) to treat HI brain injury in the neonatal mouse. Assessing the regenerative capacity of hMSCs is crucial for translation of our knowledge to the clinic. We determined the neuroregenerative potential of hMSCs in vitro and in vivo by intranasal administration 10 d post-HI in neonatal mice. HI was induced in P9 mouse pups. 1×10(6) or 2×10(6) hMSCs were administered intranasally 10 d post-HI. Motor behavior and lesion volume were measured 28 d post-HI. The in vitro capacity of hMSCs to induce differentiation of mouse neural stem cell (mNSC) was determined using a transwell co-culture differentiation assay. To determine which chemotactic factors may play a role in mediating migration of MSCs to the lesion, we performed a PCR array on 84 chemotactic factors 10 days following sham-operation, and at 10 and 17 days post-HI. Our results show that 2×10(6) hMSCs decrease lesion volume, improve motor behavior, and reduce scar formation and microglia activity. Moreover, we demonstrate that the differentiation assay reflects the neuroregenerative potential of hMSCs in vivo, as hMSCs induce mNSCs to differentiate into neurons in vitro. We also provide evidence that the chemotactic factor CXCL10 may play an important role in hMSC migration to the lesion site. This is suggested by our finding that CXCL10 is significantly upregulated at 10 days following HI, but not at 17 days after HI, a time when MSCs no longer reach the lesion when given intranasally. The results described in this work also tempt us to contemplate hMSCs not only as a potential treatment option for neonatal encephalopathy, but also for a plethora of degenerative and traumatic injuries of the nervous system.

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PKH-26 labeled hMSCs migrate to the lesion site.1×106 hMSCs were labeled with PKH-26 and administered intranasally at 10 days after HI. (A+B) Mice were terminated 24 hours after hMSC treatment. (A) PKH-26+ hMSCs in the ipsilateral damaged cortex. (B) Contralateral cortex shows no PKH-26+ signal. (C) qPCR validation of PCR array confirmed the upregulation of five genes 10 days after HI. At 17 days after HI 2 genes were down-regulated in comparison to 10 days following HI. (HI n = 10; sham n = 6). (D) hMSCs express CXCR3, which increases after co-culture with HI brain extract (n = 2). Data represent mean ± S.E.M. *p<0.05; **p<0.01; ***p<0.001 by ANOVA and Bonferroni post-hoc test. Dashed line  =  lesion border; Asterisk  =  lesion site. Blue  =  Dapi staining. Scale bar  = 50 µm.
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pone-0112339-g003: PKH-26 labeled hMSCs migrate to the lesion site.1×106 hMSCs were labeled with PKH-26 and administered intranasally at 10 days after HI. (A+B) Mice were terminated 24 hours after hMSC treatment. (A) PKH-26+ hMSCs in the ipsilateral damaged cortex. (B) Contralateral cortex shows no PKH-26+ signal. (C) qPCR validation of PCR array confirmed the upregulation of five genes 10 days after HI. At 17 days after HI 2 genes were down-regulated in comparison to 10 days following HI. (HI n = 10; sham n = 6). (D) hMSCs express CXCR3, which increases after co-culture with HI brain extract (n = 2). Data represent mean ± S.E.M. *p<0.05; **p<0.01; ***p<0.001 by ANOVA and Bonferroni post-hoc test. Dashed line  =  lesion border; Asterisk  =  lesion site. Blue  =  Dapi staining. Scale bar  = 50 µm.

Mentions: In a previous study we showed that PKH-26-labeled C57BL/6 MSCs migrate specifically towards the HI-damaged brain region within 24 hours after intranasal administration [11]. We demonstrated that the PKH-26+ signal is specific and not due to auto-fluorescence of damaged tissue as we did not see any positive signal in the HI-damaged brain after vehicle-treatment. As we are applying human MSCs to the mouse brain, we first established if intranasal delivery is an efficient administration route before investigating the therapeutic potential of hMSCs in vivo. We administered 1×106 PKH-26-labeled hMSCs intranasally at 10 days after HI and sacrificed the mice 24 hours later. At 10 days after HI, i.e. the time-point when hMSCs are administered intranasally, the entire hippocampus and part of the cortex had degenerated and an evident cyst could be discerned in the ipsilateral hemisphere. 24 hours after hMSC administration we observed a strong PKH-26+ signal, i.e. hMSCs, in the sensorimotor and epithalamic regions of the damaged brain, surrounding the cyst (Fig. 3A). We did not detect any PKH-26+ signal in the contralateral hemisphere (Fig. 3B).


Intranasal administration of human MSC for ischemic brain injury in the mouse: in vitro and in vivo neuroregenerative functions.

Donega V, Nijboer CH, Braccioli L, Slaper-Cortenbach I, Kavelaars A, van Bel F, Heijnen CJ - PLoS ONE (2014)

PKH-26 labeled hMSCs migrate to the lesion site.1×106 hMSCs were labeled with PKH-26 and administered intranasally at 10 days after HI. (A+B) Mice were terminated 24 hours after hMSC treatment. (A) PKH-26+ hMSCs in the ipsilateral damaged cortex. (B) Contralateral cortex shows no PKH-26+ signal. (C) qPCR validation of PCR array confirmed the upregulation of five genes 10 days after HI. At 17 days after HI 2 genes were down-regulated in comparison to 10 days following HI. (HI n = 10; sham n = 6). (D) hMSCs express CXCR3, which increases after co-culture with HI brain extract (n = 2). Data represent mean ± S.E.M. *p<0.05; **p<0.01; ***p<0.001 by ANOVA and Bonferroni post-hoc test. Dashed line  =  lesion border; Asterisk  =  lesion site. Blue  =  Dapi staining. Scale bar  = 50 µm.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4232359&req=5

pone-0112339-g003: PKH-26 labeled hMSCs migrate to the lesion site.1×106 hMSCs were labeled with PKH-26 and administered intranasally at 10 days after HI. (A+B) Mice were terminated 24 hours after hMSC treatment. (A) PKH-26+ hMSCs in the ipsilateral damaged cortex. (B) Contralateral cortex shows no PKH-26+ signal. (C) qPCR validation of PCR array confirmed the upregulation of five genes 10 days after HI. At 17 days after HI 2 genes were down-regulated in comparison to 10 days following HI. (HI n = 10; sham n = 6). (D) hMSCs express CXCR3, which increases after co-culture with HI brain extract (n = 2). Data represent mean ± S.E.M. *p<0.05; **p<0.01; ***p<0.001 by ANOVA and Bonferroni post-hoc test. Dashed line  =  lesion border; Asterisk  =  lesion site. Blue  =  Dapi staining. Scale bar  = 50 µm.
Mentions: In a previous study we showed that PKH-26-labeled C57BL/6 MSCs migrate specifically towards the HI-damaged brain region within 24 hours after intranasal administration [11]. We demonstrated that the PKH-26+ signal is specific and not due to auto-fluorescence of damaged tissue as we did not see any positive signal in the HI-damaged brain after vehicle-treatment. As we are applying human MSCs to the mouse brain, we first established if intranasal delivery is an efficient administration route before investigating the therapeutic potential of hMSCs in vivo. We administered 1×106 PKH-26-labeled hMSCs intranasally at 10 days after HI and sacrificed the mice 24 hours later. At 10 days after HI, i.e. the time-point when hMSCs are administered intranasally, the entire hippocampus and part of the cortex had degenerated and an evident cyst could be discerned in the ipsilateral hemisphere. 24 hours after hMSC administration we observed a strong PKH-26+ signal, i.e. hMSCs, in the sensorimotor and epithalamic regions of the damaged brain, surrounding the cyst (Fig. 3A). We did not detect any PKH-26+ signal in the contralateral hemisphere (Fig. 3B).

Bottom Line: Our results show that 2×10(6) hMSCs decrease lesion volume, improve motor behavior, and reduce scar formation and microglia activity.This is suggested by our finding that CXCL10 is significantly upregulated at 10 days following HI, but not at 17 days after HI, a time when MSCs no longer reach the lesion when given intranasally.The results described in this work also tempt us to contemplate hMSCs not only as a potential treatment option for neonatal encephalopathy, but also for a plethora of degenerative and traumatic injuries of the nervous system.

View Article: PubMed Central - PubMed

Affiliation: Lab. of Neuroimmunology and Developmental Origins of Disease, University Medical Center Utrecht, Utrecht, The Netherlands.

ABSTRACT
Intranasal treatment with C57BL/6 MSCs reduces lesion volume and improves motor and cognitive behavior in the neonatal hypoxic-ischemic (HI) mouse model. In this study, we investigated the potential of human MSCs (hMSCs) to treat HI brain injury in the neonatal mouse. Assessing the regenerative capacity of hMSCs is crucial for translation of our knowledge to the clinic. We determined the neuroregenerative potential of hMSCs in vitro and in vivo by intranasal administration 10 d post-HI in neonatal mice. HI was induced in P9 mouse pups. 1×10(6) or 2×10(6) hMSCs were administered intranasally 10 d post-HI. Motor behavior and lesion volume were measured 28 d post-HI. The in vitro capacity of hMSCs to induce differentiation of mouse neural stem cell (mNSC) was determined using a transwell co-culture differentiation assay. To determine which chemotactic factors may play a role in mediating migration of MSCs to the lesion, we performed a PCR array on 84 chemotactic factors 10 days following sham-operation, and at 10 and 17 days post-HI. Our results show that 2×10(6) hMSCs decrease lesion volume, improve motor behavior, and reduce scar formation and microglia activity. Moreover, we demonstrate that the differentiation assay reflects the neuroregenerative potential of hMSCs in vivo, as hMSCs induce mNSCs to differentiate into neurons in vitro. We also provide evidence that the chemotactic factor CXCL10 may play an important role in hMSC migration to the lesion site. This is suggested by our finding that CXCL10 is significantly upregulated at 10 days following HI, but not at 17 days after HI, a time when MSCs no longer reach the lesion when given intranasally. The results described in this work also tempt us to contemplate hMSCs not only as a potential treatment option for neonatal encephalopathy, but also for a plethora of degenerative and traumatic injuries of the nervous system.

Show MeSH
Related in: MedlinePlus