Consistent with the putative roles of CaMid1p and Cch1p as Ca2+ channels, Ca2+ accumulation in the Camid1Δ and Cacch1Δ mutants was significantly reduced after 2 hr culture in Ca2+-depleted medium supplemented with Ca2+ (p = <0.001; Supplemental Data).
Early cultivation in medium with supplemental TGF-β1/FGF-2 doubled cell fractions in 2-week constructs compared to unsupplemented controls.
When necessary the medium was supplemented with serum (or Bovine Serum Albumin, BSA), glucose, glutamine and Non Essential Aminoacids (NEAA, formulation of the 50X solution in supplemental table 2).
Briefly, cells were plated onto specialty plates (Costar ultra low attachment clusters), containing the culture medium lacking supplemental LIF.
After transduction, 3 × 10 cells were cultured on MS5 stromal cells in Long-Term Culture medium (see Supplemental Experimental Procedures).
Plant were allowed to acclimatize for two days after transplanting and the soil was then saturated with the modified MS medium without or with supplemental 100 mM NaCl, as required.
Bovine calf articular chondrocytes were seeded onto biodegradable polyglycolic acid scaffolds and cultured for four weeks using in vitro systems providing different mechanical environments (static and mixed Petri dishes, static and mixed flasks, and rotating vessels) and different biochemical environments (medium with and without supplemental insulin-like growth factor I, IGF-I).
Water and swab specimens were plated onto buffered charcoal yeast extract medium with and without supplemental antimicrobial drugs; standard plating techniques were used.
In complete medium with a low supplemental level of VEGFA (∼2 ng/ml), VEGFA or CRIM1 knockdown HUVECs exhibited slower growth kinetics (Fig. 7A), as shown previously (Fig. 6A).
Then the medium was replaced with a supplemental medium containing 10% FBS, and the cells were cultivated for 24-96 h under standard conditions.
