Adult neural stem and progenitor cells modified to secrete GDNF can protect, migrate and integrate after intracerebral transplantation in rats with transient forebrain ischemia

Abstract

Adult neural stem and progenitor cells (NSPCs) are important autologous transplantation tools in regenerative medicine, as they can secrete factors that protect the ischemic brain. We investigated whether adult NSPCs genetically modified to secrete more glial cell line-derived neurotrophic factor (GDNF) could protect against transient ischemia in rats. NSPCs were harvested from the subventricular zone of adult Wistar rats and cultured for 3 weeks in the presence of epidermal growth factor. The NSPCs were treated with fibre-mutant Arg-Gly-Asp adenovirus containing the GDNF gene (NSPC-GDNF) or enhanced green fluorescent protein (EGFP) gene (NSPC-EGFP; control group). In one experiment, cultured cells were transplanted into the right ischemic boundary zone of Wistar rat brains. One week later, animals underwent 90 min of intraluminal right middle cerebral artery occlusion followed by magnetic resonance imaging and behavioural tests. The NSPC-GDNF group had higher behavioural scores and lesser infarct volume than did controls at 1, 7 and 28 days postocclusion. In the second experiment, we transplanted NSPCs 3 h after ischemic insult. Compared to controls, rats receiving NSPC-GDNF had decreased infarct volume and better behavioural assessments at 7 days post-transplant. Animals were killed on day 7 and brains were collected for GDNF ELISA and morphological assessment. Compared to controls, more GDNF was secreted, more NSPC-GDNF cells migrated toward the ischemic core and more NSPC-GDNF cells expressed immature neuronal marker. Moreover, the NSPC-GDNF group showed more effective inhibition of microglial invasion and apoptosis. These findings suggest that NSPC-GDNF may be useful in treatment of cerebral ischemia.

Rescue of radiation-induced cognitive impairment through cranial transplantation of human embryonic stem cells

“…Whether those cells were actually integrated into hippocampal circuits is not yet known. Regarding trophic support, studies of ischemic injury found transplanted neural stem/precursor cells to be neuroprotective via glial cell line-derived neurotrophic factor (GDNF) (48,49). Other work using a transgenic model of Alzheimer disease provided evidence that brain-derived neurotrophic factor (BDNF) from transplanted rodent neural stem cells (mNSC) played a significant role in restoring certain cognitive deficits when assessed 1-month posttransplantation (31).…”

Regenerative Medicine for Cerebral Infarction

“…In experiments with animals, many researchers have confirmed that stem cell transplantation provides neuroprotective effects immediately after transplantation, based on behavioral analyses and histological analyses that show reductions in infarct volume (Kameda et al, 2007;Kurozumi et al, 2004;Takahashi et al, 2008). Histological analyses also showed that the neuroprotective effects were due to enhanced angiogenesis (Onda et al, 2008), anti-apoptotic effects (Kameda et al, 2007;Kurozumi et al, 2004), and so on.…”

“…Regarding iPS cells, for example, Maekawa et al showed that using maternal transcription factor Glis1 instead of oncogenic c-Myc enhanced the generation of iPS cells when expressed together with key transcription factors Oct3/4, Sox2, and Klf4 (Maekawa et al, 2011). The majority of animal experiments have demonstrated the neuroprotective effect of transplantation using allografts of adult NSCs or MSCs in the acute phase of ischemia (Kameda et al, 2007;Takahashi et al, 2008), but the effectiveness of stem cell transplantation during the subacute or chronic phase of ischemia was not seen (de Vasconcelos Dos Santos., 2011). To avoid the problem of rejection by the immune system, and ethical issues, autologous stem cell transplantation using adult NSCs and MSCs is attractive, but considerable time is needed to expand these cells so that sufficient quantities are available.…”

“…Currently, many different types of stem cell can be cultured and transplanted, including induced pluripotent stem cells (iPS cells) (Takahashi & Yamanaka, 2006), embryonic stem cells (ES cells) (Wang et al, 2011), neural stem cells (NSCs) (Kameda et al, 2007;Muraoka et al, 2006) , mesenchymal stem cells (MSCs) (Kurozumi et al, 2004;Wang et al, 2010) , and hematopoietic stem cells (Shyu et al, 2006), and some of these have already been used in clinical trials. Every type of stem cell has unique advantages and disadvantages.…”

“…Stem cell transplantation is one of the most widely employed strategies using exogenous stem cells. Many studies with experimental animals have shown that stem cell transplantation enhances functional recovery after cerebral infarction (Kameda et al, 2007;Takahashi et al, 2008). Based on the results of animal experiments, several clinical trials for patients with cerebral infarction are currently ongoing, using stem cell transplantation techniques, typically with mesenchymal stem cells (Detante et al).…”