title: Methylcellulose during cryopreservation of ventral mesencephalic tissue fragments fails to improve survival and function of cell suspension grafts.
abstract: Cryopreservation may allow long-term storage of fetal ventral mesencephalon (VM) for transplantation in patients suffering from Parkinson's disease (PD). We investigated whether the polymer methylcellulose protects fetal rat VM during cryopreservation in liquid nitrogen and improves survival and function of this tissue as intrastriatal suspension grafts in the 6-hydroxydopamine (6-OHDA) rat model. VM tissue fragments (E14-E15) were either immediately dissociated and grafted as a cell suspension (FRESH) or cryopreserved under controlled conditions for 7 days in a conventional cryoprotective medium (CRYO) or a medium containing 0.1% methylcellulose (mCRYO) and then dissociated and grafted. Rats from the cryo-groups showed only limited behavioral compensation in contrast to complete compensation observed in rats from the FRESH group. Cryopreservation of fetal rat VM decreased the viability of cell suspensions in vitro to about 70%, survival of grafted tyrosine hydroxylase-immunoreactive (TH-IR) neurons to 11% and 20%, and transplant volume to 8% and 17% (mCRYO and CRYO, respectively, compared to FRESH). The addition of 0.1% methylcellulose to tissue fragments during freezing did neither improve in vitro viability nor survival of TH-IR neurons nor behavioral compensation when compared to the control CRYO group. These results suggest that methylcellulose failed to improve survival of cryopreserved dopaminergic ventral mesencephalic neurons.
authors.affiliation: Klinikum Grosshadern, Department of Neurology, Ludwig-Maximilians-University, Munich, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Sautter
authors.forename: S
authors.initials: S
authors.lastname: Strecker
authors.forename: A
authors.initials: A
authors.lastname: Kupsch
authors.forename: W H
authors.initials: WH
authors.lastname: Oertel
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D017690:Cell Transplantation; D003342:Corpus Striatum; D015925:Cryopreservation; D005260:Female; D016332:Fetal Tissue Transplantation; D006085:Graft Survival / Q000187:drug effects; D008636:Mesencephalon / Q000187:drug effects* / Q000196:embryology / Q000637:transplantation; D008747:Methylcellulose / Q000494:pharmacology*; D009043:Motor Activity / Q000187:drug effects; D009498:Neurotoxins; D016627:Oxidopamine; D051381:Rats; D017207:Rats, Sprague-Dawley; D012399:Rotation
chemical_list: D009498:Neurotoxins; D016627:Oxidopamine; D008747:Methylcellulose