Evaluation of characteristic of human turbinate derived mesenchymal stem cells cultured in the serum free media

Abstract

We evaluated the effect of serum-free and xeno-cultivation (SFXFM) on the characterization, proliferation, and differentiation properties of human nasal stem cells (airway tissue; hTMSCs). hTMSCs were isolated from 10 patients, after which patient samples were separated into two groups, an SFXFM group and a control group. The control group was treated with bovine serum-containing medium. FACS analysis revealed that SFXFM-cultured hTMSCs maintained a characteristic mesenchymal stem cell phenotype. hTMSC proliferation was not influenced by SFXFM. In addition, upregulation of IL-8 and GM-CSF and downregulation of RANTES expression were shown in response to SFXFM. Moreover, two-lineage differentiation properties (osteocyte and adipocyte) of hTMSCs were enhanced under SFXFM. Finally, the genetic stability of SFXFM-cultured hTMSCs was demonstrated by normal karyotype results. SFXFM enables good expansion, multipotentiality, and normal genotype maintenance of MSCs. Moreover, this approach serves as a substitute to conventional media for the cultivation of capable MSCs for upcoming medical applications.

Genetic Stability of Mesenchymal Stromal Cells for Regenerative Medicine Applications: A Fundamental Biosafety Aspect

“…A high number of studies have well documented that human MSCs acquire cytogenetic alterations in culture both in autosomes and in sex chromosomes, even if they are frequently transient and not clonal [28,29,55,73,74,130,143,144,145,146,147,148,149,150,151,152,153]. Conversely, other studies reported stable chromosomes in culture for MSCs of different tissue origin [24,33,36,48,116,118,120,124,125,126,127,149,153,154,155,156,157,158]. Depending on the studies, tissue sources, culture conditions, culture times, these alterations occur at early or late passages and at different frequencies, thus explaining inconsistencies in literature data (for a detailed list of described aneuploidies, see Rebuzzini et al [148]).…”

“…Finally, culture conditions can greatly influence the biological properties of MSCs [50,118,154,158,174] including genomic stability [148]. Supplements seem to not influence genomic stability [36], whereas oxygen tension is critical [175] exerting a marked regulatory effect on cell cycle checkpoint genes [166]. It was demonstrated that long-term cultures under hypoxic conditions prevent senescence [82,83,84] while maintaining MSC differentiation potential.…”

“…The most commonly used MSC sources are bone marrow (BM) [17] and adipose tissue (AT) [18,19,20,21]. They can also be isolated from a series of other tissues like umbilical cord (UC) or Wharton’s Jelly [22], cord blood [23,24], placenta and amnion [25,26], amniotic fluid [27], chorionic villi [28], dental pulp [29,30], endometrium [31], brain [32], skin [33], synovial membrane and synovial fluid [15,34], sweat glands [35], nasal mucosa [36].…”

Acceleration of Translational Mesenchymal Stromal Cell Therapy Through Consistent Quality GMP Manufacturing

“…For identity characterization, using gene expression assays such as Taqman TM to replace standard Tri-lineage differentiation staining assay is another example. While the current Tri-lineage differentiation assay requires 2–3 weeks for the cells to differentiate followed by staining and imaging ( Owston et al, 2019 ; Sanjurjo-Rodriguez et al, 2019 ; Rajpar and Barrett, 2020 ), newer methods such as the measurement of hMSC Tri-lineage gene expression following 1–2 weeks of differentiation reduces the time needed while providing quantitative information relative to the current method ( Szepesi et al, 2016 ; Hwang et al, 2017 ; Ling et al, 2020 ).…”