Technical library
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The Chondrocyte Growth Medium SupplementMix consists of Fetal Calf Serum (final concentration 10 % v/v) which has specifically been tested for the culture of primary chondrocytes. FCS contains a variety of different growth factors, which are however, not analyzed in more detail. There are no further growth factors added to the SupplementMix.
The optimal calcium concentration for both proliferation and differentiation of keratinocytes depends on the species and also on the media formulation. To keep primary human keratinocytes in the proliferative status, concentrations between 0.03 and 0.15 mM (PromoCell Keratinocyte Growth Medium 2: 0.06 mM) are generally used. Increasing the calcium above 1 mM will induce terminal differentiation and lead to the loss of proliferative activity.
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.
CD146 (MUC18) is a surface marker, expressed on pericytes, MSCs, and endothelial cells from large vessels (but not microvascular endothelial cells). In combination with CD34 (a marker for endothelial and hematopoietic cells), pericytes can be characterized by FACS analysis as CD146+/CD34-. The absence of CD34 expression makes sure that the cells are not of hematopoietic/endothelial origin. Further markers that have been described for pericytes are NG2, CD90, alpha-SMA, and PDGFR-beta.
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.
Our HPF (C-12360) are isolated from peripheral lung tissue.
Recommended procedure: - Thaw the vial according to our protocol - Seed one part of the cells (5,000 cells/cm²) directly into multiwell plates for subsequent differentiation induction; grow in Preadipocyte Growth Medium until they reach 100% confluency, then induce differentiation according to the recommended protocol - Seed the other part (5,000 cells/cm²) into a TC dish and expand them in Preadipocyte Growth Medium; trypsinize at subconfluence; if necessary divide the cell suspension again into two parts and use one part for differentiation tests (see above), the other part to continue the culture of undifferentiated HWP. - Differentiation capacity may decline after 1-2 passages in vitro, therefore best perform your differentiation tests at early passages. HWP at higher passages can still be used for studies that require undifferentiated HWP (e.g. proliferation assays).
The basic information we receive from the surgeons about the tissue donors usually includes age, gender, and ethnicity.
- For many of our donors of subcutaneous preadipocytes, we also have information on BMI, hair color, skin pigmentation, and, in some cases, smoking habits or known diseases (e.g. Diabetes). Most of our subcutaneous HWP donors are between ∼20-65 years old.
- The visceral HWP donors are mostly between ∼20-85 years old. For many cell lots we know the BMI, in some cases also the hair color, skin pigmentation, smoking habits, and/or known diseases (e.g. Diabetes or COPD).
If you are looking for particular specifications, please contact our Scientific Support, so that we can offer you appropriate HWP lots.
Further Information
We guarantee 15 population doublings for our NHEM. In terms of passages this corresponds to approx. 6-8 subcultivations.
We can however not guarantee that the activity of all melanocyte genes remains unchanged during this time. Primary cells do gradually change their phenotype in vitro. Therefore, it is recommended to use the cells for the important experiments at low passages.
Mature chondrocytes from articular cartilage are postmitotic producers of extracellular matrix components like collagen types II, IX, and XI and proteoglycans. Upon release from the tissue of origin and seeding in monolayer culture, cells re-enter the cell cycle and proliferate. After a period of approx. 1-3 weeks, they will gradually start dedifferentiation and will adopt a fibroblastic phenotype. Dedifferentiation can be detected by decreased collagen type II or increased collagen type I expression, as well as by the appearance of the fibroblast marker Thy-1/CD90. Re-differentiation can be induced by changing from conventional monolayer culture to a 3-D culture system.
All our chondrocytes (C-12710) are isolated either from knee joint or femoral head. If you need chondrocytes specifically from either localization, please contact our Technical Customer Service before placing your order. They will send you the available lot numbers from the localization that you are looking for.
We source the bronchial tissue from forensic medicine and from thoracic surgery. For some lots, the smoking habits of the donors are known. Please contact our Technical Customer Service before ordering the cells if you need this information.