Cryopreserved single donor primary HBMEC isolated from normal human brain were purchased from Cell Systems (Kirkland, WA) at passage 2. Each preparation was extensively characterized as previously described [31].
In the simulations performed here, we investigated the impact of a one-time use (i.e. during a single generation) of cryopreserved semen.
Cryopreserved single-donor primary human brain microvascular endothelial cells (HBMEC) isolated from normal human brain and reported to be mycoplasma negative by the provider were purchased from Cell Systems Corp. (Kirkland, WA, USA) at passage 2. Preparations were analysed from nine different donors.
Thaw-and-use target cells in our experiments were pre-labeled with the fluorescent dye calcein AM, cryopreserved in single-use aliquots and used directly in assays after thawing.
Cells were passaged every 10 14 days into single cell suspensions before being cryopreserved.
We also applied the technology to determine chromosome copy number on 134 single-blastomeres derived from 26 cryopreserved embryos from six unrelated couples.
NHBE cells (Lonza, Walkersville, MD, USA) from a single healthy male donor were expanded, cryopreserved and maintained in culture in an air/liquid interface system, as previously described.
Most parasitic protozoa respond to cryopreservation stresses similarly to other single cell suspensions, but few species are currently routinely cryopreserved by protocols specifically designed for vitrification.
To report a live birth after the transfer of a single blastocyst derived from a human oocyte cryopreserved for 5 years.
Cryopreserved human hepatocytes from a single donor were seeded within the HμREL® device to predict the in vivo hepatic clearance (CLH) of six marketed model compounds (carbamazepine, caffeine, timolol, sildenafil, imipramine, and buspirone).
