JOURNAL OF TRANSLATIONAL MEDICINE
PUBBLICATO IL 27 GENNAIO 2015
Human neural stem cell transplantation in ALS: initial results from a phase I trial.
Mazzini L 1, Gelati M 2,3, Profico DC 4,5, Sgaravizzi G 6, Projetti Pensi M 7,8, Muzi G 9, Ricciolini C 10,11, Rota Nodari L 12,13, Carletti S 14, Giorgi C 15, Spera C 16, Domenico F 17, Bersano E 18, Petruzzelli F 19, Cisari C 20, Maglione A 21, Sarnelli MF 22, Stecco A 23, Querin G 24, Masiero S 25, Cantello R 26, Ferrari D 27, Zalfa C 28, Binda E 29,30, Visioli A 31, Trombetta D 32, Novelli A 33, Torres B 34, Bernardini L 35, Carriero A 36, Prandi P 37, Servo S 38, Cerino A 39, Cima V 40, Gaiani A 41, Nasuelli N 42, Massara M 43, Glass J 44, Sorarù G 45, Boulis NM 46, Vescovi AL 47,48,49,50.
We report the initial results from a phase I clinical trial for ALS. We transplanted GMP-grade, fetal human neural stem cells from natural in utero death (hNSCs) into the anterior horns of the spinal cord to test for the safety of both cells and neurosurgical procedures in these patients. The trial was approved by the Istituto Superiore di Sanità and the competent Ethics Committees and was monitored by an external Safety Board.
Six non-ambulatory patients were treated. Three of them received 3 unilateral hNSCs microinjections into the lumbar cord tract, while the remaining ones received bilateral microinjections. None manifested severe adverse events related to the treatment, even though nearly 5 times more cells were injected in the patients receiving bilateral implants and a much milder immune-suppression regimen was used as compared to previous trials.
No increase of disease progression due to the treatment was observed for up to18 months after surgery. Rather, two patients showed a transitory improvement of the subscore ambulation on the ALS-FRS-R scale (from 1 to 2). A third patient showed improvement of the MRC score for tibialis anterior, which persisted for as long as 7 months. The latter and two additional patients refused PEG and invasive ventilation and died 8 months after surgery due to the progression of respiratory failure. The autopsies confirmed that this was related to the evolution of the disease.
We describe a safe cell therapy approach that will allow for the treatment of larger pools of patients for later-phase ALS clinical trials, while warranting good reproducibility. These can now be carried out under more standardized conditions, based on a more homogenous repertoire of clinical grade hNSCs. The use of brain tissue from natural miscarriages eliminates the ethical concerns that may arise from the use of fetal material.
TRIAL REGISTRATION: EudraCT:2009-014484-39 .
NEURAL PROGENITOR CELLS
Methods in Molecular Biology pp 65-77
PUBBLICAZIONE 15 LUGLIO 2013
Culturing and expansion of "clinical grade" precursors cells from the fetal human central nervous system.
Gelati M 1, Profico D, Projetti-Pensi M, Muzi G, Sgaravizzi G, Vescovi AL.
NSCs have been demonstrated to be very useful in grafts into the mammalian central nervous system to investigate the exploitation of NSC for the therapy of neurodegenerative disorders in animal models of neurodegenerative diseases. To push cell therapy in CNS on stage of clinical application, it is necessary to establish a continuous and standardized, clinical grade (i.e., produced following the good manufacturing practice guidelines) human neural stem cell lines. In this chapter, we illustrate some of the protocols routinely used into our GMP cell bank for the production of "clinical grade" human neural stem cell lines.
Neural Stem Cell Assays
PUBBLICAZIONE 15 LUGLIO 2013
Cryopreservation of Human Neural Stem and Progenitor Cells
Daniela Celeste Profico 1, Giada Sgaravizzi1, Massimo Projetti Pensi 1, Gianmarco Muzi 1, Claudia Ricciolini 1, Maurizio Gelati 1,2 and Angelo Luigi Vescovi 1,2
Neural stem cells (NSCs) are, by definition, capable of unlimited self-renewal in vitro; nevertheless, there are limitations in the number of passages to avoid cell senescence and minimize the possibility of karyotype alterations. For this reason NSC cryopreservation seems to be fundamental to maintain NSC functionality in transplant experiments or to use NSCs in further derivation of neural lineages. Here, we describe a useful and detailed freezing method for NSCs.