Our kidney cells are characterized by their epithelial cell morphology and by cytokeratin expression using a pan-cytokeratin antibody.

Our human pulmonary microvascular endothelial cells (C-12281) are sourced from the lung parenchyma with all large vessels being removed beforehand. Therefore most of the HPMEC originate from capillaries.

Our HPF (C-12360) are isolated from peripheral lung tissue.

All cytokines in Cytokine Mix E (human TPO SCF flt-3 ligand and IL-3) are produced in E.coli. They are purified by chromatography are free of endotoxins and are tested for their biological activity.

Alcian Blue stains the extracellular matrix of chondrocytes; e.g. cartilage-specific aggrecan and other glycosaminoglycans. To our knowledge; chondrocytes only express aggrecans when grown in 3-D culture and not in 2-D culture. To detect cartilage specific markers in monolayer culture; it is recommended to perform immunofluorescence detection of collagen type II.

Human Mesenchymal Stem Cells have retained the ability to differentiate into a variety of cell types including fat cells; chondrocytes; and osteoblasts. PromoCell supply hMSC from 3 different tissues: bone marrow (hMSC-BM; C-12974); umbilical cord matrix (hMSC-UC; C-12971); and adipose tissue (hMSC-AT; C-12977).

MSC-BM show very good differentiation into bone cells but also into chondrocytes and fat cells when the respective Differentiation Media is used.
MSC-UC have a high potential to differentiate into chondrocytes but only weak potential for fat or bone cell differentiation.
In contrast; MSC-AT differentiate very well into fat and bone cells but only moderately into chondrocytes.

In other words; to obtain a high percentage of bone cells; MSC-BM or MSC-AT are the cells of choice. There are of course lot-to-lot variations and the differentiation will decrease from passage to passage. If you need MSCs with a particularly high osteoblast differentiation capacity; you can call our Technical Customer Service before placing your order so that we can select an appropriate cell lot for you.

Adipogenic differentiation can be identified morphologically and without any staining by the formation of intracellular lipid vesicles.
In contrast; when MSC differentiate into bone cells; there is no significant change in morphology. It is recommended to perform Alkaline Phosphatase staining to detect osteoblastic differentiation or Alizarin Red S staining to show osteoblast mineralization.
Chondrogenic differentiation is generally performed as spheroids in 3-D cell culture and not in 2-D monolayer culture. Staining with Alcian Blue to visualize the differentiation process is indispensable.
Neurogenic differentiation can be detected using neuron specific markers (e.g. beta-3 tubulin; NeuN; MAP2) and by their typical neuronal morphology.

For differentiation protocols; please see attached Application Notes.

Our hMSC-AT are characterized by their differentiation potential into chondrocytes; fat cells; and bone cells. In addition; we determine the presence of CD73; CD90 and CD105 expression as well as the absence of CD14; CD19; CD34; CD45 & HLA-DR expression by flow cytometry as proposed by the International Society for Cellular Therapy.

Mononuclear cells are mostly used in immunology; infection biology; hematology and cancer research to study subpopulations of blood cells. Our Mononuclear Cell Medium (C-28030) is intended for short-term maintenance (up to 48 hrs) of the thawed hMNC before you proceed with your experiments. The number of PDs will depend on the subsequent cell culture conditions and is not determined by PromoCell. Please note: Depending on the conditions; the ratio of the subpopulations will gradually change; as the different blood cell types behave in different manners. Researchers normally start soon after thawing to either select the cell type of their interest (e.g. hematopoietic cells; endothelial progenitor cells) or perform experiments with all populations of hMNCs (e.g. to study effects on toxicity; viability or metabolism).

We have tested the differentiation capacity of our hMSC into adipocytes chondrocytes and osteoblasts over time and still see good differentiation rates after 10 population doublings i.e. at passage 5. However the differentiation potential declines with ongoing population doublings. To obtain optimal differentiation rates experiments should be performed as early in culture as possible.

For chrondrogenic differentiation it is important that the cells do not adhere to the wells. During differentiation the cells form spheroids which float in the medium. Therefore there are no special requirements for the wells as long as they are U bottom shaped and suitable for suspension culture.

The following criteria are applied for blood donations: a) Donors with light infection like a cold or cough are excluded for one week b) Donors with infections with a temperature above 37.9°C and/ or antibiotics therapy are excluded for 4 weeks c) Blood pressure: exclusion only if systolic blood pressure is < 100 mm Hg or > 180 mm Hg or if diastolic blood pressure is > 100 mm Hg. No exclusion if blood pressure is drug treated and in acceptable range. d) High cholesterol: no exclusion even though medication is used e) Diabetes: exclusion if diabetes type I or use of insulin f) Steroid use: exclusion for 4 weeks after application g) Cancer: exclusion h) Other chronic diseases: exclusion depends on the type of disease (for example chronic heart disease; autoimmune disease)

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