What is a CFC assay?

The colony forming cell (CFC) assay is used to study the proliferation and differentiation pattern of hematopoietic progenitors by their ability to form colonies in a semisolid medium.

How do you do a colony formation assay?

A clonogenic assay, also known as a colony formation assay In order to measure clonogenicity, cells need to be seeded at very low densities and left for a period of 1-3 weeks for colonies to form. Colonies are then fixed, stained with crystal violet to make them visible, and counted.

What is colony forming cell?

The colony forming cell (CFC) assay, also referred to as the methylcellulose assay, is an in vitro assay used in the study of hematopoietic stem cells. The assay is based on the ability of hematopoietic progenitors to proliferate and differentiate into colonies in a semi-solid media in response to cytokine stimulation.

What is MethoCult?

MethoCult™ H4434 Classic (MethoCult™ GF H4434) is a complete methylcellulose-based medium for the growth and enumeration of hematopoietic progenitor cells in colony-forming unit (CFU) assays of human bone marrow, mobilized peripheral blood, peripheral blood, and cord blood samples.

How is CFU measured?

The CFU/ml can be calculated using the formula: cfu/ml = (no. of colonies x dilution factor) / volume of culture plate. For example, suppose the plate of the 10^6 dilution yielded a count of 130 colonies.

What does CD34 stand for?

CD34 is a marker of human HSC, and all colony-forming activity of human bone marrow (BM) cells is found in the CD34+ fraction. Clinical transplantation studies that used enriched CD34+ BM cells indicated the presence of HSC with long-term BM reconstitutional ability within this fraction.

What does 1 CFU mean?

cfu stands for colony-forming unit. This means that cfu/g is colony-forming unit per gram and cfu/ml is colony-forming unit per millilitre. A colony-forming unit is where a colony of microbes grow on a petri dish, from one single microbe.

What is CFU count?

A colony-forming unit (CFU, cfu, Cfu) is a unit used in microbiology. It estimates the number of bacteria or fungal cells in a sample which are viable, able to multiply via binary fission under the controlled conditions.

How are hematopoietic stem cells cultured?

A) Human CD34+ hematopoietic stem cells isolated from normal, healthy human bone marrow were cultured in either Stemline™ HSC media in methylcellulose with Epo or competitor media in methylcellulose with Epo. Cultures were scored after 12 – 16 days of culture using an inverted microscope.

How many cells are in a CFU?

While doing this you are assuming that one cell will form one colony. But you don’t know, may be 2 or 3 cells form one colony. Since you are not sure than you express the number as colony forming units or cfu per ml. the forming unit can be one cell or more.

What is CD34-positive mean?

In normal conditions, CD34-positive cells account for about 1–2% of the total bone marrow cells. The TdT+ subset of precursor B cells (hematogones) is also positive for CD34. Approximately 40% of AMLs and over 50% of ALLs express CD34. CD34 is also expressed in a garden variety of nonhematopoietic tumors.

What is CD34 marker used for?

CD34 is routinely used to identify and isolate human hematopoietic stem/progenitor cells (HSPCs) for use clinically in bone marrow transplantation, but its function on these cells remains elusive.

How do you calculate coliform count?

Use the upper ideal range (60 for fecal coliforms) with the smallest volume (0.1 mL) and calculate the result: (60 ÷ 0.1) x 100 = >60,000 CFU/100 mL. When MF counts are above and below the reporting range, select the most nearly acceptable count to the ideal range.

How do you calculate CFU?

1. To find out the number of CFU/ ml in the original sample, the number of colony forming units on the countable plate is multiplied by 1/FDF. This takes into account all of the dilution of the original sample.
2. 200 CFU x 1/1/4000 = 200 CFU x 4000 = 800000 CFU/ml = 8 x 10.
3. CFU/ml in the original sample.

Why CFU mL is important?

The CFU method has two noteworthy advantages, namely the capacity for counts of any number of bacteria using dilutions, if too many, or concentrations if too few. Second, only viable bacteria are counted with this method as the CFU method excludes dead bacteria and debris.

What is the difference between HSC and Hspc?

HSC are defined by their ability to self-renew and to produce all blood cell types, while HPC do not have self-renewal capacity and are more restricted in the mature blood cells they can produce. Investigators typically use a combination of functional and phenotypic characteristics to categorize populations of HSPC.