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As previously published [23], for every 105 cells adoptively transferred, approximately 10% were CD45hi and 5% were CD45hiCD8+.
Bombarded globular soybean somatic embryos (SSE) that were cultured on D20 were transferred (approximately 100 clumps of 0.3-0.4 0.3-0.4eter per plate) onto selecmion mediametertaining 5 mM AEC.
Given that the average hyphal strand is 12 μm wide, and using the calculated nuclear densities (see below), we estimated that our asexual propagation regimen transferred approximately 25 million nuclei between successive culture tubes.
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Different components as PEG 1000 g ∙ mol− 1, Na2SO4, H3PO4, phosphoric and acid impurities as Al III), Ca II), Fe III), Cd II), V(IV) were titrated in each phase of the systems showing that it was possible to transfer approximately a 75% of loaded H3PO4 to the polymer rich phase.
A 96-pin (2 mm diameter) pin tool (V&P Scientific, Inc., San Diego, CA, USA) was used, transferring approximately 3 µl of cell culture at each passage.
Slotted metal pins (946MP6B, Arrayit) were used to transfer approximately 2.4 nL of monomer solution onto 10 pHEMA-coated substrates before slides were irradiated with a long wave UV source for 30 s.
In this case, the ratio of data transferred is approximately equal to the ratio of their mean residence times.
We then transferred spheroids approximately 300 µm in diameter from free suspension into 1% agarose gel and cultured them under three conditions: normal medium without external compression, starvation medium (no glucose, no serum and 1% oxygen) without compression or normal medium with external compression (see experimental setup in Supplementary Fig. S1C).
A total of 13 15 replicates per each MNIL were set up, each consisting of a group of 20 transferred eggs (approximately 21 000 transferred eggs in total).
For vitrification, each blastocyst was first incubated in holding medium (TCM-Hepes supplemented with 20% cattle serum) containing 7.5% ethylene glycol and 7.5% dimethyl sulphoxide for 3 min, and then transferred in approximately 1 to 2 μl of solution to a 20- μl droplet holding medium containing 16.5% ethylene glycol, 16.5% dimethyl sulphoxide and 0.5 M sucrose.
For total heat transfer, Al2O3 gives somewhat higher heat transfer (approximately 5%) when compared to the CuO nanofluid.
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