Kontur P J, Leranth C, Redmond D E, Roth R H, Robbins R J
Yale University Neural Transplant Program, Department of Pharmacology, Yale University School of Medicine, New Haven, Connecticut 06510.
Exp Neurol. 1993 Jun;121(2):172-80. doi: 10.1006/exnr.1993.1084.
Both fresh and cryopreserved-thawed human fetal ventral mesencephalon have been used for preclinical research and implantation into the brains of patients with Parkinson's disease. Further characterization and an evaluation of the effects of cryopreservation on immunocytochemical and neurochemical markers of monoamine neurons in human fetal ventral mesencephalic tissue are reported here. Fresh and cryopreserved-thawed human fetal mesencephalic tissue of 7-10 weeks fetal age was analyzed for the presence of tyrosine hydroxylase-like immunoreactivity and levels of dopamine (DA), norepinephrine, serotonin, 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), and 5-hydroxyindole acetic acid. After fixation, cryopreserved-thawed mesencephalic tissue exhibited cellular tyrosine hydroxylase-like immunoreactivity identical to that seen in fresh tissue. The levels of DA, norepinephrine, serotonin, and 5-hydroxyindole acetic acid in the cryopreserved-thawed tissue were the same as the levels in fresh tissue. The levels of DOPAC were higher, and those of HVA were lower, in the cryopreserved-thawed tissue compared to the levels in fresh tissue. The changes in the levels of the DA metabolites, DOPAC and HVA, without corresponding change in the levels of the parent monoamine in cryopreserved-thawed tissue, indicate ongoing metabolic activity in DA-containing neurons. These results further suggest that cryopreservation and subsequent thawing does not have a measurable adverse effect on DA biosynthesis in the human fetal mesencephalon. The presence of the monoamines and their metabolites in the ventral mesencephalon at 7-10 weeks of fetal age coincides well with the early presence of immunocytochemical markers of monoamine neurons and reflects the early function of the nuclear groups containing specific monoamine neurotransmitters.
新鲜的和冷冻解冻后的人类胎儿腹侧中脑均已用于临床前研究以及植入帕金森病患者的大脑。本文报告了对冷冻保存对人类胎儿腹侧中脑组织中免疫细胞化学和单胺能神经元神经化学标志物影响的进一步表征和评估。分析了胎龄7 - 10周的新鲜和冷冻解冻后的人类胎儿中脑组织中酪氨酸羟化酶样免疫反应性以及多巴胺（DA）、去甲肾上腺素、血清素、3,4 - 二羟基苯乙酸（DOPAC）、高香草酸（HVA）和5 - 羟吲哚乙酸的水平。固定后，冷冻解冻后的中脑组织呈现出与新鲜组织中相同的细胞酪氨酸羟化酶样免疫反应性。冷冻解冻后组织中DA、去甲肾上腺素、血清素和5 - 羟吲哚乙酸的水平与新鲜组织中的水平相同。与新鲜组织相比，冷冻解冻后组织中DOPAC的水平较高，而HVA的水平较低。冷冻解冻后组织中DA代谢物DOPAC和HVA水平的变化，而母体单胺水平没有相应变化，表明含DA神经元中存在持续的代谢活性。这些结果进一步表明，冷冻保存和随后的解冻对人类胎儿中脑的DA生物合成没有可测量的不利影响。胎龄7 - 10周时腹侧中脑中单胺及其代谢物的存在与单胺能神经元免疫细胞化学标志物的早期存在非常吻合，并反映了含有特定单胺神经递质的核团的早期功能。
