|LETTER TO EDITOR
|Ahead of print publication
The need for recreating rheumatic disease biology in a dish
Sakir Ahmed1, Soumya R Mohapatra2
1 Department of Research and Development; Department of Clinical Immunology and Rheumatology, Kalinga Institute of Medical Sciences, Kalinga Institute of Industrial Technology, Deemed to be University, Bhubaneswar, Odisha, India
2 Department of Research and Development, Kalinga Institute of Medical Sciences, Kalinga Institute of Industrial Technology, Deemed to be University; School of Biotechnology, Kalinga Institute of Industrial Technology, Deemed to be University, Bhubaneswar, Odisha, India
|Date of Submission||28-Oct-2021|
|Date of Acceptance||29-Oct-2021|
|Date of Web Publication||28-Mar-2022|
Soumya R Mohapatra,
Department of Research and Development, Kalinga Institute of Medical Sciences, Kalinga Institute of Industrial Technology, Deemed to be University, Bhubaneswar, Odisha
Source of Support: None, Conflict of Interest: None
We have read with great interest the protocol for the development of indigenous primary cell cultures from different sources by Sharma et al. The strengths of the protocol are simplicity and cost-effectiveness. The advent of cell-culturing techniques revolutionized the field of biomedical sciences and contributed immensely to the advancement of drug development and medicine. Due to the ease of maintaining them in culture, well-established and immortalized cell lines form the foundation of the current world of biomedical research. However, the rampant use of these established cell lines comes with their own unique set of problematic issues, which needs addressing. The most common among these issues is the deviation in biological behavior and fundamental characteristics of these established immortal cells from the physiological cell types from which they have been derived., These deviations are resultant of changes in environmental stimuli and/or accumulation of genetic changes over a long course of time while the cells are in continuous culture in vitro.
To overcome the accumulation of genetic changes, standard practice among biomedical researchers is to restrict the number of passages an established cell line is subcultured. This holds across different disease spectra ranging from cell lines for glioblastomas to osteogenic and adipogenic cells to fibroblasts from rheumatoid arthritis (RA). Even for most well-established immortalized cell lines, standard practice is to restrict the number of subculture passages at 30. When this passage number is achieved, researchers usually discard these older cells in favor of low passage number cells of the same cell line by thawing and subculturing new vials of cells taken out from long-term storage in liquid nitrogen. Despite these drawbacks, the convenience provided by the in vitro culturing of immortal cells has assured their continued relevance in the biomedical research arena.
The continued desire of clinical researchers to closely mimic physiological processes in vitro, however, restricts the universal use of immortalized cell lines. Instead, this necessitates the establishment of primary cultures with cells derived by biopsies from the immediate site of origin, whose physiological processes need to be recreated. Primary cell cultures facilitate the near-exact replication of biological processes found in vivo, thus reducing and/or eliminating the ethical dilemma associated with using various animal models and human studies. The necessity for primary cell cultures is of utmost relevance for rheumatic diseases, as most animal models currently available fail to recapitulate the human disease to the fullest extent. This could be attributed to the fact that most of these animal models are incapable of developing autoimmune diseases naturally, thus necessitating the artificial induction of autoimmunity in these model systems.
For any new drug development, the first step is always tested in animal models and in cell lines. Having fresh patient-derived cells lines enables the investigator to mimic in vivo conditions as closely as possible. The use of fibroblast-like synoviocytes (FLSs) cultures has enabled us to learn so much about how FLSs behave in RA as induced aggressors having malignancy cell-like invasive potential. One step further would be to have three-dimensional (3-D)-printed scaffolding for the growth of the primary cell lines. A 3-D model system with cocultured FLSs and macrophage-like synoviocytes with interspersed endothelial cells with vascular architecture would be the most ideal model of synovial tissue.
Thus, the simplified protocol for a primary fibroblast culture can enable upcoming scientists to develop their own culture facilities to work with patient material, instead of depending on commercial available cell lines deprived of patients with entirely different disease states. Moreover, the potential is almost endless.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Sharma S, Pande G, Rai M, Agarwal V. Indigenous primary culture protocols for human adult skin fibroblast, pancreatic stellate cells, and peritoneal fibroblasts. Indian J Rheumatol 2021;16:298. [Full text]
Neumann E, Riepl B, Knedla A, Lefèvre S, Tarner IH, Grifka J, et al.
Cell culture and passaging alters gene expression pattern and proliferation rate in rheumatoid arthritis synovial fibroblasts. Arthritis Res Ther 2010;12:R83.
Zeng Y, Wang X, Wang J, Yi R, Long H, Zhou M, et al.
The tumorgenicity of glioblastoma cell line U87MG decreased during serial in vitro
passage. Cell Mol Neurobiol 2018;38:1245-52.
Kwist K, Bridges WC, Burg KJ. The effect of cell passage number on osteogenic and adipogenic characteristics of D1 cells. Cytotechnology 2016;68:1661-7.
Richter M, Piwocka O, Musielak M, Piotrowski I, Suchorska WM, Trzeciak T. From donor to the lab: A fascinating journey of primary cell lines. Front Cell Dev Biol 2021;9:711381.
Damerau A, Gaber T. Modeling rheumatoid arthritis in vitro
: From experimental feasibility to physiological proximity. Int J Mol Sci 2020;21:7916.
Bottini N, Firestein GS. Duality of fibroblast-like synoviocytes in RA: Passive responders and imprinted aggressors. Nat Rev Rheumatol 2013;9:24-33.