The fluidlab R-300 enables the analysis of diverse cell types, including fibroblasts. These cells are found throughout the body and play a major role in wound healing, tumor growth and skin rejuvenation. Their research enables new therapeutic options against cancer, but also the development of new anti-aging procedures. The range of applications is wide and therefore research into cells is of particular interest.
Learn more in this article about
- what fibroblasts are,
- how fibroblasts are structured,
- where fibroblasts occur,
- the importance of fibroblasts in research,
- how fibroblast cells can be verified with the fluidlab R300 and
- how cell validation of fibroblasts using the fluidlab is also possible in animals.
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What are fibroblasts?
Fibroblasts are division-active cells of the connective tissue. The cells derive from the mesoderm and are not fully differentiated. Therefore, they can differentiate into various connective tissue cells, such as the immobile form, fibrocytes. They also form all the important components of connective tissue - including collagen - and thus determine the properties of the tissue. Both the regeneration and the remodeling of connective tissue originate from them.1
Myofibroblasts are a special form of fibroblasts
Myofibroblasts were discovered in experimental studies on inflammation and wound healing. They have a fine structural similarity to smooth muscle cells in that they contain microfibrils, microtendons and cell-cell channels.
This special type is a modified form of fibroblasts, which are capable of contracting. Characteristically, myofibroblasts express alpha-smooth muscle actin.
Cytokines serve as mediators that control the activation and modification of fibroblasts to myofibroblasts. The cells have been detected in human granulation tissue, palmar fibromatosis and reactive stroma in invasive carcinomas.
Under normal conditions, they are also located
- in the skin,
- at the hair follicle,
- in the periodontal ligament and
- in lung septa.
Myofibroblasts are responsible for wound closure. During inflammation - or wound healing - there is proliferation of fibroblasts and subsequent modulation of these into myofibroblasts. Collagen type 3 is also modulated.
The next step in wound healing is the transition to fibrocytes and collagen type 1. Finally, contraction and wound closure occur.2 Two central roles are fulfilled by myofibroblasts.
On the one hand, they produce a very large amount of collagen.3 This characteristic helps in the closure of wounds, because the collagen forms a net-like layer over the injured area, which serves as a temporary wound closure. On the other hand, the contraction of myofibroblasts, which are connected to the wound edges and to each other via fibronectin filaments, further supports wound closure.4
Low level production of the cells inhibits wound healing, whereas overproduction of this form of fibroblast, is characteristic in tumor-like processes.5 In other vessels, such as testicular tubules and capillaries, as well as mucous membranes, myofibroblasts are responsible for maintaining tension and volume.
The structure of fibroblasts
Fibroblasts have an elongated shape. At their tips, they have a large number of projections which allow them to contact each other. The nucleus of fibroblasts is round to oval and the nucleolus is strongly defined. They have many cell organelles. These are responsible for the formation of matrix components. Their cytoplasm is basophilic and the chromatin, also called euchromatin, is very fine.6
Where do fibroblasts occur?
Primarily, fibroblasts are specific resident cells of connective tissue. Connective tissue is found throughout the body and provides support, strength and elasticity to organs. Fibroblasts are found in the interstitial spaces of the organs.
Fibroblasts are also found in the lung. There, they are located in a highly complex, multicellular environment, which is normally closely adjacent to the epithelium or endothelium.7
However, fibroblasts can also emerge from differentiated connective tissue cells, such as bone or cartilage. This occurs in response to external stimuli, for example injury.8
Can fibroblasts be grown outside the organism?
After the fibroblasts have been taken from a cell association or organism, they can be grown in vitro. They grow solitarily and not in tissue. In addition, they are not fully differentiated, which makes them easy to maintain in culture. Their proliferation is very rapid.
They can be grown as a primary cell line. In this case, their lifespan is limited to approximately 50 division cycles. However, it is also possible to convert them into permanent cell lines by genetic intervention.9
This ability to grow fibroblasts in culture vessels for laboratory studies makes them attractive for experimentation.
The importance of fibroblasts in research
Fibroblasts play a particularly important role in wound healing and regeneration of the skin. For this reason, they are especially important in the development of new therapeutic options that affect the skin. Diseases of the connective tissue, pulmonary fibrosis or even tumors can thus be better understood by studying fibroblasts.
Fibroblasts also play a special role in the development of angiogenesis models. Angiogenesis is the formation of new blood vessels from existing ones. In this process, endothelial cells migrate to the point where the blood vessels are to be formed and form into thin capillaries.
In vitro angiogenesis models use fibroblasts that secrete growth factors, which in turn initiate the formation of blood vessels by endothelial cells.10 These in vitro models can then be used to test the influence of various substances or drugs on the vasculature. In addition, it can be used to supply in vitro organs.
Fibroblasts are of particular interest in the aesthetic field. There they are said to serve the rejuvenation of the skin.
Investigations into the use of fibroblasts in anti-aging
Fibroblasts are the main cells of connective tissue. There they are very important for strength, as they produce supporting molecules, among other things. In the skin, they support the absorption of moisture. That is why research into anti-aging products and treatments also focuses on fibroblasts.
The collagen produced by the fibroblasts provides increased strength of the extracellular matrix. With age, the number of fibroblasts in the connective tissue decreases. They become less productive and the tissue loses strength. Regenerative technologies can be used to act directly on the cells to counteract this. The skin's own fibroblasts can be injected into the skin for this purpose.
After about 25-30 years, the skin begins to age biologically. Collagen, hyaluronic acid and elastin, which are among the most important components of the intercellular matrix, decrease. As a result, the skin becomes thinner, elasticity and firmness decrease, and wrinkles appear and become deeper. This is caused by the 10-15% decrease in fibroblasts every 8-10 years.
This can be counteracted by withdrawing fibroblasts at a young age and cryopreserving them. In this way, they can be stored for a longer period without losing their vitality when they are thawed later.
When the thawed fibroblasts are inserted into the skin, they are able to synthesize the whole set of components of the intercellular matrix. Consequently, the skin looks better, it gains firmness and elasticity. The aging process is slowed down as a result.
This type of anti-aging procedure is natural in the sense that it occurs naturally and simply utilizes one's biological reserves.11
Analysis of fibroblasts provides important insights for cancer research
A 2017 study found that fibroblasts reduce the effectiveness of chemotherapy for pancreatic cancer.12 Patients with this type of advanced cancer are typically treated with a chemotherapy drug called gemcitabine. However, the drug prolongs survival by only a few months. Yet the effect of gemcitabine is promising in test trials with pancreatic cancer cells in the Petri dish.
The chemotherapeutic agent is added to the cells or the organism in its inactive form and is only converted into an active form there. Enzymes attach phosphate groups to the molecule and thus catalyze it into gemcitabine triphosphate, the biologically active form. This is incorporated into DNA as a building block. During DNA synthesis, it thus prevents cell division.
Now it turns out that the connective tissue cells surrounding the tumors take up the therapeutic agent and metabolize it into the active form. However, it cannot pass through the cell membrane of the fibroblasts. For this reason, it cannot reach the tumor cells in the body and act there.
This knowledge about fibroblasts enables research into a better method of combating metastases. However, this is also proving difficult, since, for example, therapy methods that also attack the connective tissue around the tumor could lead to new metastases in the organism. Experiments with genetically modified mice showed that the drug initially worked better in animals with less connective tissue, but that they died earlier because more new metastases formed in them.13
However, a study from 2015 proved that connective tissue slows down the growth of tumors. This was demonstrated by removing fibroblasts from experimental animals with tumors in the pancreas. Consequently, the tumor grew faster. According to this, fibroblasts around the tumor ensured that it was encapsulated so that it could not be further supplied with nutrients.14
Fibroblasts can lead to tumors due to their increased growth. These can be benign fibromas as well as malignant fibrosarcomas.
Fibromas can occur on any part of the body that contains connective tissue. They can occur in soft and hard form. The formation of fibromas occurs due to the increased proliferation of fibroblasts. This results in increased tissue growth in one location. What causes the increased growth is unknown. However, this type of tumor is not dangerous and remains so.15
This is different in the case of fibrosarcoma. These malignant growths are also found in connective tissue, mostly on the legs. Rarely, it also occurs on the arms. The exact causes of formation are also unknown here. However, they are caused by fibroblasts that divide at an above-average rate.16
In summary, fibroblasts play an important role in tumor growth, whether they surround it or lead to its formation themselves through their increased proliferation. For these reasons, they are of high interest in cancer research and may help in the study of their behavior to new therapeutic methods
This is how the verification of fibroblast cells works with the fluidlab R300
The fluidlab R-300 can be used to count fibroblasts and measure their viability. No dye is used for this, which makes the measurement faster. In addition, the viability of the fibroblasts is not affected in this way.
The fluidlab R-300 is the first portable automated cell counter. While other automated cell counters are often bulky and for this reason bound to a fixed place in the laboratory, the fluidlab R-300 is many times smaller and lighter.
This offers the possibility to perform cell counts at different working locations and can be used, for example, directly under the sterile workbench to save space.
In addition, it offers other advantages:
- The holographic microscope does not need to be calibrated.
- Due to the automatic focus, the focusing plane does not have to be selected by the user.
- The neural network counts the fibroblasts reliably with a high statistical certainty.
- A small sample volume is already sufficient to obtain reliable results.
This facilitates and speeds up the counting, as well as the analysis of fibroblasts in the laboratory.
You would like to make use of the advantages of the fluidlab R-300 yourself? Feel free to contact our customer service at anvajo.contact@gmail.com.
How fast can the fluidlab detect and count fibroblasts?
The holographic microscope of the fluidlab R-300 creates holograms of the fibroblasts, which contain valuable information about the sample. The captured images are analyzed by machine learning algorithms to determine the cell number and viability of the fibroblasts. The result of the measurement is then available within 30 seconds.
Which cell sizes can be analyzed with the fluidlab?
The fluidlab is able to count cells from 3 µm - 80 µm. Viability is recorded for cells with a size of 8 µm - 80 µm. Since fibroblasts have a size of up to approx. 50 µm, they can be analyzed by the fluidlab without any problems.
Cell validation of fibroblasts is also possible in animals using the fluidlab R-300
Animal cells do not differ in structure from human cells. Both cell types are surrounded by a selectively permeable plasma membrane. The cytoplasm is located inside and within it the nucleus as well as the cell organelles.17
For this reason, it is possible to validate not only human but also animal fibroblasts with the fluidlab R-300. This can be particularly important for cats, as they frequently develop fibrosarcomas, malignant mesenchymal tumors. These tumors arising from fibroblasts are consequences of spontaneous mutations, the formation of which need not have any particular cause.
However, a clustered occurrence of these tumors has been noted in cats, especially after vaccination.18 The fluidlab R-300 also allows validation of fibroblasts in this case.
1 Sauermost R. et al., 1999, Fibroblasten. Lexikon der Biologie, Spektrum Akademischer Verlag, https://www.spektrum.de/lexikon/biologie/fibroblasten/24391 [28.05.20]
2 Meister, P., 1998, Myofibroblasten Übersicht und Ausblick. Pathologe 19, 187–193
3 Nonnenmacher, 2019, Myofibroblasten, https://medlexi.de/Myofibroblasten [28.05.20]
4 Vgl. Nonnenmacher, 2019
5 Vgl. Meister, P., 1998
6 Schöni-Affolter F., Das Bindegewebe. Division of Histology, https://www3.unifr.ch/apps/med/elearning/de/bindegewebe/downloads/bg_print_d.pdf [04.06.20]
7 Lynne A. et al., 2009, Fibroblasts. Asthma and COPD (Second Edition), https://www.sciencedirect.com/topics/neuroscience/fibroblast [05.06.20]
8 Vgl. Sauermost R., et al., 1999
9 Vgl. Sauermost R. et al., 1999
10 Andrew C. Newman et al., 2011, The requirement for fibroblasts in angiogenesis: fibroblast-derived matrix proteins are essential for endothelial cell lumen formation. Molecular Biology of the Cell Vol. 22, No. 20, https://doi.org/10.1091/mbc.e11-05-0393 [11.09.20]
11 2017, https://www.cosmacon.de/glossary/fibroblasten/ [11.09.20]
12 Hessmann et al. 2017 Fibroblast drug scavenging increases intratumoural gemcitabine accumulation in murine pancreas cancer, Gut. 2018 Mar;67(3):497-507
13 Der seltsame Appetit der Fibroblasten https://www.doccheck.com/de/detail/articles/612-der-seltsame-appetit-der-fibroblasten [01.09.20]
14 2015, Wie die Forschung versucht den Krebs in Schach zu halten, Focus, https://www.focus.de/gesundheit/ratgeber/krebs/therapie/neue-therapieansaetze-krebs-keine-lokal-begrenzte-erkrankung_id_5003398.html [01.09.20]
15 Mai, Manuela 2019, Fibrom: Entfernung nur aus kosmetischen Gründen notwendig. https://www.focus.de/gesundheit/ratgeber/krebs/therapie/neue-therapieansaetze-krebs-keine-lokal-begrenzte-erkrankung_id_5003398.html [01.09.20]
16 Fibrosarkom- Ursachen, Symptome & Behandlung https://medlexi.de/Fibrosarkom [01.09.20]
17 Minuth W. et al., 2003, Zukunftstechnologie Tissue Engineering: Von der Zellbiologie zum künstlichen Gewebe, Zellen und Gewebe, Regensburg, WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim, S. 5
18 https://www.zooplus.de/magazin/katze/katzengesundheit-pflege/fibrosarkom [11.09.20]
