Technical Library
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NHEK.f are isolated from tissue samples (foreskin) of single donors; aged between 1-10 years. NHEK.f pooled are prepared from the foreskins of 3 individual donors. The cells of each donor are expanded in separate TC vessels and the cells are pooled after secondary culture; before cryopreservation.
Both NHEK from single donor and NHEK pooled are in P2 after thawing.
Differentiation of hMSC into mature adipocytes takes approx. 2 weeks. You can keep the adipocytes for up to 3 weeks in the MSC Adipogenic Differentiation Medium. After 3 weeks we recommend to switch to PromoCell's Adipocyte Nutrition Medium (C-27438).
Most of our subcutaneous preadipocyte lots achieve > 80-90% differentiation when differentiation is induced at P2 (directly after thawing).
We generally recommend using cells for differentiation tests that haven't undergone more than 4-5 doublings (a maximum of 1 passage after thawing); as the differentiation ratio will decline with the age of the cells.
The source is porcine pancreas.
The source of our heparin is ex porcine mucosa.
Our Human Renal Cortical Epithelial Cells (C-12660) are isolated from the cortex of the human kidney. The renal cortex is the outer portion of the kidney. It contains the renal corpuscles the proximal and distal convoluted tubules and the cortical collecting ducts.
The cells sink down to the bottom of the culture vessel but don't really attach. They retain a roundish morphology and can be rinsed off with culture medium easily.
Our HREpC (C-12665) comprise a heterogenous mixture of renal epithelial cells isolated from cortex and medulla. The HRCEpC (C-12660) are derived from renal cortex only.
The population doubling times for PromoCell hMSC (hMSC-BM; hMSC-AT; hMSC-UC) are typically ≤ 30 hrs when using Mesenchymal Stem Cell Growth Medium 2 (C-28009). If you seed the hMSC at 4.000 cells/cm2; it will take between 4-7 days until they reach subconfluency.
PromoCell's Normal Human White Preadipocytes (HWP) are isolated from adult subcutaneous or visceral adipose tissue from different locations. The cells are frozen in our serum-free freezing medium (Cryo-SFM) at the end of passage 1 (= secondary culture). A randomly selected vial is then used for quality control; which includes determination of growth characteristics; control of morphology; and tests for differentiation capacity into mature adipocytes.
The recommended seeding density of preadipocytes after thawing/trypsinization is 5;000 cells/cm2; cells should be trypsinized before reaching 90% confluence. Population doubling times are usually between 20-50 hrs (10 population doublings guaranteed). Using a 1:4 split ratio; you can perform ∼4-5 passages with the cells.
Preadipocyte Growth Medium (C-27417) is used to propagate the cells. To induce differentiation of preadipocytes into adipocytes; cells are grown in PromoCell's Preadipocyte Growth Medium until they reach 100% confluency. Cells are then cultured in Preadipocyte Differentiation Medium (C-27437) for 72 h; followed by 10-14 days in Adipocyte Nutrition Medium (C-27439). During this time the cells start to accumulate fat droplets which can be visualized under the microscope.
We recommend performing differentiation experiments at population doubling numbers lower than 4-5; in order to reach a high differentiation level of the culture.
No; we don't perform CD90 immunomagnetic separation with our pericytes. We can however exclude fibroblast contamination as follows:
1) The presence of fibroblasts in our pericyte cultures would be detectable shortly after cell isolation; as pericytes need up to 2 weeks before they start proliferating. Fibroblasts on the other hand would proliferate immediately and overgrow the culture.
2) Characterization of the isolated pericytes during QC includes flow cytometry of CD146. Pericytes express CD146 whereas placental fibroblasts don't.
Our hPC-PL (C-12980) are isolated from microvessels of the human placenta; from the chorionic villi.
The number of populations doublings is not determined for each individual cell lot; but in our experience; they can be grown for at least 15 population doublings.