Technical Library
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PromoCell Preadipocytes are isolated from subcutaneous or visceral fat.
Subcutaneous fat is found just underneath the skin and it is not related to the obesity-linked diseases. Its accumulation represents the normal physiological buffer for excess energy intake. When the storage capacity of subcutaneous fat is exceeded or the generation of new adipocytes is impaired; visceral fat starts to accumulate. It is located in the abdomen and around internal organs (e.g. kidney; heart; or bladder) and it is linked to hypertension; diabetes; and cardiovascular disease.
Adipocytes from subcutaneous and visceral fat differ in several functions; like their response to insulin and other hormones or their lipolytic activity. It highly depends on the scientific problem being addressed; which of the two cell types are best suited for your experiments.
FCS is a natural product consisting of many different components like salts hormones vitamins trace elements proteins and enzymes. There can be large lot-to-lot variations between different serum batches regarding the concentrations of growth-promoting factors. The higher the serum content in a culture medium the higher the impact of the variations on the cell culture system. For this reason PromoCell has developed several serum-reduced and serum-free media where part of the serum has been replaced by more defined factors like cytokines hormones or vitamins.Media with reduced serum concentrations have the benefit that they produce more standardized culture conditions over a long time span.
Please find attached a trouble shooting guide to identify possible reasons for poor detachment during subculture.
Poor attachment after thawing can be a result of inappropriate freezing storing or thawing the cells as well as from inadequate culture conditions (medium incubator). The attached trouble shooting guide should help you to identify the possible reasons.
Yes it is possible to transfect our normal human cells. In general primary and normal cells are much harder to transfect than immortalized cell lines. Apart from the cell type successful transfection also depends on the culture's age and density at transfection the vector used the purity of the nucleic acids the composition of the transfection medium and the experimental conditions.
Both trypsin and accutase represent mixtures of different proteolytic enzymes. Trypsin is prepared from porcine pancreas accutase from invertebrates. Accutase can replace trypsin for the detachment and dissociation of anchorage-dependent cells from surfaces and can also be used on suspension cells to reduce clumping in preparation for counting. The advantages of accutase over the traditional trypsin treatment are that it is more gentle and less damaging to cells (leading to increased viability) and does not contain any mammalian or bacterially derived proteins. Accutase is more thermolabile than trypsin and usually doesn't require an inactivation step.
For the expansion of hematopoietic progenitors (CD133+ cells; CD34+ cells); PromoCell has developed Hematopoietic Progenitor Cell Expansion Medium XF (C-28021); a serum-free and xeno-free formulation. The medium must be supplemented with either Cytokine Mix E (C-39890) or with the user's own mixture of cytokines. Cytokine Mix E is a ready-to-use mix containing recombinant human TPO; SCF; flt3-ligand; and IL-3.
The strong expansion of the progenitor cells in Expansion Medium XF + Cytokine Mix E persists for at least two weeks; resulting in a 200-300x increase of the cell number with only 20-30% differentiated cells.
Note: If starting with CD133+ cells; the CD133 marker is getting lost during this expansion step. The resulting cells are D34+/CD38-/CD133-.
Our HAoAF (C-12380) are isolated from the Adventitia the outer layer of the aorta. The cells have been characterized as fibroblasts by the expression of fibroblast-specific CD90.
The PromoCell quality control procedure includes the cultivation of fibroblasts for 15 population doublings. This is to ensure that the cells can be grown for a minimum of 15 PDs (6-8 passages depending on the split ratio used) but it does not mean that the cells immediately senesce after that point. We don't determine the maximum number of doublings or passages but most NHDF lots will certainly achieve > 20 passages.
HFDPC are isolated from the hair papilla of normal human scalp hair follicles. Hair papilla in the adult hair follicle play a crucial role in the dermal-epidermal interactions that control hair production and in hair growth cycle events. The follicle dermal cells are cryopreserved at second passage and can be cultured for at least 10 population doublings when using PromoCell Follicle Dermal Papilla Cell Growth Medium (Cat. C-26501). Typical population doubling times are between 20-36 hrs. The recommended seeding density after thawing/trypsinization is 5;000-10;000 cells/cm2. Using 1:4 splits; you can perform 4-5 passages with the cells.
PromoCell's Osteoblast Basal Medium is an optimized media formulation developed for human osteoblast culture. The exact composition is proprietary.
Our subcutaneous HWP are isolated from subcutaneous fat of different localizations; e.g. abdomen; breast; or upper arm.
The visceral preadipocytes are isolated from fat surrounding e.g. the pericardium; or from the omentum or mediastinum.
The exact localization is specified in the Certificate of Analysis. If you need HWP from a particular localization; please contact our Technical Customer Support prior to placing your order.