BD PureCoat cultureware

Increased cell attachment and differentiation of RCG cells on BD PureCoat amine

A-C. RCG cells were freshly isolated from rat brains and seeded onto 24-well tissue culture (TC)-treated, Competitor C, or BD PureCoat amine plates in medium containing 10% serum, glutamine, and potassium chloride. After 20-22 hours, cultures were observed under a microscope and images captured at 200X magnification. Results from a representative experiment show that RCG cells attach and differentiate on BD PureCoat amine, whereas those on TC-treated and Competitor C plates are poorly attached and formed clumps.

D. Neurite outgrowth was observed from RCG cells plated on BD PureCoat amine. In some instances, cultures were incubated for 48 hours, fixed, and immunostained with anti-tubulin IIIb. RCG cells grown on BD PureCoat amine exhibit dendritic processes and express tubulin IIIb, a neuronal marker. RCG cells cultured on TC-treated and Competitor C plates remained unattached and were washed off the plates during the fixation protocol (data not shown).

Enhanced cell growth of BHK-21 cells in reduced serum on BD PureCoat amine

A. BHK-21 cells were seeded onto 96-well black/clear TC-treated, Competitor C, or BD PureCoat amine plates at densities between 1600-6400 cells/well and grown in culture medium containing 5% tryptose and 1% fetal bovine serum for 72 hours. MTS reagent (Promega) was added directly into culture wells (n=8 wells/condition) and incubated for 2 hours. Representative relative absorbance units (mean ± SEM) from one of three experiments is shown. Increased cell proliferation was observed on BD PureCoat amine at all three seeding densities tested compared to TC-treated or Competitor C plates.

B. Shown is a representative image of BHK-21 cells grown on 24-well TC-treated or BD PureCoat amine plates and fixed with 4% paraformaldehyde. A nuclear stain, DAPI, was added to cultures and fluorescence images were captured at 200X magnification. An increase in DAPI staining was observed on BD PureCoat amine indicating more cells attach and proliferate on the BD PureCoat amine surface versus TC-treated.

Improved post-wash adherence of EcoPack2-293 cells (a transformed HEK-293 cell line) to BD PureCoat amine

A. Ten thousand cells were seeded onto TC-treated, Competitor C, or 384-well black/clear BD PureCoat amine plates and grown under serum-free conditions for 20-24 hours. The cells were then washed (on an Embla cell washer) two times with Hank’s buffered salt solution containing 10 mM Hepes, loaded with calcein AM for 1 hour and read on an EnVision® (PerkinElmer) plate reader. Before and after wash images indicate that cells remain attached on BD PureCoat amine surface and are washed off the other surfaces that were tested.

B. Accordingly, relative fluorescent unit values are higher on BD PureCoat amine (data not shown). Intraplate CV values (n=240 wells) are below 10% on BD PureCoat amine offering superior consistency in cell-based assays, whereas CV values on TC-treated or Competitor C plates are much greater.

76% more Baby Hamster Kidney (BHK-21) cell growth in reduced serum on BD PureCoat amine

BHK-21 cells were seeded onto 75 cm² area flasks of TC-treated and BD PureCoat amine surface at 1.125 × 10⁶ cells/flask in 15 ml of GMEM culture medium containing 5% tryptose broth and 1% fetal bovine serum. After 72 hours, cells were trypsinized using 0.25% trypsin-EDTA for 3-5 minutes at 37°C (3 ml/flask) and the resulting suspension was centrifuged and enumerated. Results from a representative experiment show ≥ 76% increase in BHK-21 growth on BD PureCoat amine vs. TC (n=3 flasks/surface).

Up to 140% more human bone-marrow derived mesenchymal stem cells (hMSCs) on 6-well BD PureCoat amine surface.

Cryopreserved hMSCs (P2) were thawed and immediately seeded onto 6-well TC-treated (TC) or BD PureCoat amine plates in serum-containing MSCGM™ media (Lonza) at ~ 6,000 cells/cm². After 4 days of growth, these cells were trypsinized, and split at a 1:2 ratio. Cells were grown for an additional 3 days, re-passaged (P3) and split in 1:5 ratio for two successive passages (P4 and P5) each grown for ~ 4 days. An increased number of cells are obtained more rapidly on BD PureCoat amine as compared to a TC-treated surface with increases ranging from 25-140% across several passages (P3-P5). This growth advantage starts slow but gains momentum by P3 and is continued through P5.

Up to 102% more human adipose-derived stem cells (hASCs) on BD PureCoat amine surface.

Cryopreserved hASCs (P2) were thawed and immediately seeded onto 6-well TC-treated or BD PureCoat amine plates in serum-containing MSCGM media (Lonza) at ~6,000 cells/cm². After 4 days of growth, P2 cells were trypsinized and split at a 1:5 ratio. Cells were grown for an additional 4 days (P3) and split at a 1:4 ratio for two successive passages (P4 and P5). Shown are calculated hASC cell yields from one donor. Cells were passaged between 70-80% growth confl uency as measured by IncuCyte™ and the split ratio was kept constant across both TC-treated and BD PureCoat amine surface. A greater total cell yield of hASCs cultured on BD PureCoat amine was observed and increases in cell yield compounded with every passage. BD PureCoat amine-expanded hMSCs and hASCs retained differentiation potential.