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Neuroregenerative bystander-effects of conditioned media from adipose tissue-derived fibroblast-like cells in vitro

Pedachenko E.G., Liubich L.D., Staino L.P., Egorova D.M., Skaterna T.D.


SUMMARY. The use of different types of stem cells, including me-senchymal multipotent stromal cells (MMSCs) and neurogenic stem/progenitor cells (NSCs/NPCs), is a promising direction in the development of new technologies for the treatment of central nervous system diseases. An alternative to direct cell transplantation may be the use of their conditioned media (CM) as a source of secretome and a key component of the mechanism to realize their potential. Current clinical trials, using cell therapy in case of CNS pathology, mostly use adipose tissue, bone marrow, umbilical cord, and cord blood as a source of isolating MMSCs or mononuclear cells of the stromal-vascular fraction, and the lines of immortalized neurogenic cells, isolated from structures of the brain or spinal cord of the human embryo, as a source of NSCs/NPCs. In experimental conditions, in particular in rodents, adipose tissue is among the most available sources of allogenic progenitor cells of the mesenchymal type, and fetal brain is an accessible source of neurogenic cells. The aim was to study the neuroregenerative effects of conditioned media from rat adipose tissue-derived fibroblast-like cells and fetal neurogenic cells in vitro. CM from 24-h cultures of rat adipose tissue-derived fibroblast-like cells (ad-FLCs) and fetal neurogenic cells (NCs, E14) were examined using electrophoresis in 10% polyacrylamide gel. On day 57, in 2D cultures of rat neural cells (E14) the scratch assay was performed and nutrient medium DMEM with 10 % FCS (standard culture conditions, control) or 0.10 mg/ml (by amount of protein) ad-FLCs CM or NCs CM were added. Microscopic and morphometric studies were performed during four days of cultivation. After mechanical transection in the culture of neural cells under standard conditions from the first to the third day there were processes of endogenous regeneration, which decreased to the fourth day. The addition of ad-FLCs CM or NCs CM contributed to a significant increase in the degree and duration of endogenous regeneration processes in neural cell culture. Under the exposure to ad-FLCs CM up to the fourth day, the number of migrated cells and the density of cell processes in the scratch area increased 7 and 12.5 times, respectively; the exposure to NCs CM increased the number of migrated cells (3.5 times), the distance of cell migration (1.4 times), the density of cell processes (13 times). The length of the overgrown section of the scratch area increased 1.7 times after the exposure to ad-FLCs CM, NCs CM 3 times, amounting to 23.7 and 43.5 % respectively of the total length of the transection zone. 10 protein fractions were detected in the ad-FLCs CM: predominant 12, 15, 23, 30, 80 kDa and minor 28, 35, 55, 65, 75 kDa; in NCs CM 9 fractions: prevailing 15, 23, 30, 35 kDa and minor 37, 40, 46, 67, 80 kDa. CM from 24-h cultures of rat ad-FLCs or NCs (E14) stimulate endogenous regeneration processes in rat brain cell culture with mechanical monolayer transection. The affecting factors of neuroregenerative bystander effects of rat ad-FLCs or NCs are secreted biologically active proteins components of the CM predominant and minor protein fractions.

Key words: conditioned medium, adipose tissue-derived fibroblast-like cells, neurogenic cells, scratch assay, electrophoresis, protein fractions

Tsitologiya i Genetika 2022, vol. 56, no. 2, pp. 38-48

  1. The State Institution Romodanov Neurosurgery Institute, the National Academy of Medical Sciences of Ukraine
  2. Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine

E-mail: lyubichld

Pedachenko E.G., Liubich L.D., Staino L.P., Egorova D.M., Skaterna T.D. Neuroregenerative bystander-effects of conditioned media from adipose tissue-derived fibroblast-like cells in vitro, Tsitol Genet., 2022, vol. 56, no. 2, pp. 38-48.

In "Cytology and Genetics":
E. G. Pedachenko, L. D. Liubich, L. P. Staino, D. M. Egorova & T. D. Skaterna Neuroregenerative Bystander-Effects of Conditioned Media from Adipose Tissue-Derived Fibroblast-Like Cells in Vitro, Cytol Genet., 2022, vol. 56, no. 2, pp. 139147
DOI: 10.3103/S0095452722020098


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