Batch Effects during Human Bone Marrow Stromal Cell Propagation Prevail Donor Variation and Culture Duration: Impact on Genotype, Phenotype and Function

Brachtl, Gabriele; Poupardin, Rodolphe; Hochmann, Sarah; Raninger, Anna M.; Jürchott, Karsten; Streitz, Mathias; Schlickeiser, Stephan; Oeller, Michaela; Wolf, Martin; Schallmoser, Katharina; Volk, Hans‐Dieter; Geißler, Sven; Strunk, Dirk

doi:10.3390/cells11060946

Cited by 12 publications

(15 citation statements)

References 69 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To minimize the influence of in vitro culture conditions, similar isolation methods and identical culture conditions were chosen for all cell types as established and validated previously (20). There is substantial evidence that hPL supports culture expansion of various stromal cell types better than FBS or human serum without compromising their regenerative capacity (49)(50)(51).…”

Section: Origin Culture Identity and Purity Of Stromal Cellsmentioning

confidence: 99%

The enhancer landscape predetermines the skeletal regeneration capacity of stromal cells

Hochmann

Poupardin

et al. 2023

Sci. Transl. Med.

Self Cite

View full text Add to dashboard Cite

Multipotent stromal cells are considered attractive sources for cell therapy and tissue engineering. Despite numerous experimental and clinical studies, broad application of stromal cell therapeutics is not yet emerging. A major challenge is the functional diversity of available cell sources. Here, we investigated the regenerative potential of clinically relevant human stromal cells from bone marrow (BMSCs), white adipose tissue, and umbilical cord compared with mature chondrocytes and skin fibroblasts in vitro and in vivo. Although all stromal cell types could express transcription factors related to endochondral ossification, only BMSCs formed cartilage discs in vitro that fully regenerated critical-size femoral defects after transplantation into mice. We identified cell type–specific epigenetic landscapes as the underlying molecular mechanism controlling transcriptional stromal differentiation networks. Binding sites of commonly expressed transcription factors in the enhancer and promoter regions of ossification-related genes, including Runt and bZIP families, were accessible only in BMSCs but not in extraskeletal stromal cells. This suggests an epigenetically predetermined differentiation potential depending on cell origin that allows common transcription factors to trigger distinct organ-specific transcriptional programs, facilitating forward selection of regeneration-competent cell sources. Last, we demonstrate that viable human BMSCs initiated defect healing through the secretion of osteopontin and contributed to transient mineralized bone hard callus formation after transplantation into immunodeficient mice, which was eventually replaced by murine recipient bone during final tissue remodeling.

show abstract

Section: Origin Culture Identity and Purity Of Stromal Cellsmentioning

confidence: 99%

The enhancer landscape predetermines the skeletal regeneration capacity of stromal cells

Hochmann

Poupardin

et al. 2023

Sci. Transl. Med.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Exposure of cells to FBS during isolation and expansion steps might affect the cell phenotype and their differentiation capacity. A recent study showed that BMSCs lost their chondrogenic potential after expansion in FBS containing medium [82]. Thus, the serum substitute medium for hBMSCs isolation and expansion also needs to be optimized to have fully FBS-free studies.…”

Section: Discussionmentioning

confidence: 99%

Development of serum substitute medium for bone tissue engineering

Ansari

Ito

Hofmann

2022

Preprint

View full text Add to dashboard Cite

In tissue engineering, cells are grown often on scaffolds and subjected to chemical/mechanical stimuli. Most such cultures still use fetal bovine serum (FBS) despite its known disadvantages including ethical concerns, safety issues, and variability in composition, which greatly influences the experimental outcomes. To overcome the disadvantages of using FBS, chemically defined serum substitute medium needs to be developed. Development of such medium depends on cell type and application - which makes it impossible to define one universal serum substitute medium for all cells in any application. Here, we developed a serum substitute medium for bone tissue engineering (BTE) in a step-by-step process. Essential components were added to the medium while human bone marrow mesenchymal stromal cells (hBMSCs, osteoblast progenitor cells) were cultured in two-dimensional (2D) and three-dimensional (3D) substrates. In a 3-week culture, the developed serum substitute medium worked equally well as FBS containing medium in term of cell attachment to the substrate, cell survival, osteoblast differentiation, and deposition of extracellular matrix. In the next step, the use of serum substitute medium was evaluated when culturing cells under mechanical loading in the form of shear stress. The outcomes showed that the application of shear stress is essential to improve extracellular matrix formation while using serum substitute medium. The developed serum substitute medium could pave the way in replacing FBS for BTE studies eliminating the use of controversial FBS and providing a better-defined chemical environment for BTE studies.

show abstract

“…Afterwards the sections were washed in acetic acid n -butyl ester (P036.1 Roth) and mounted. Cartilage (proteoglycan specific) stains bright red and the backgrounds stains in green [ 47 ]. For Masson–Goldner trichrome and elastica staining, kits were used according to the manufacturer’s recommendations (Morphisto 12043, Morphisto 14604) [ 48 ].…”

Section: Methodsmentioning

confidence: 99%

Chondrocyte Isolation from Loose Bodies—An Option for Reducing Donor Site Morbidity for Autologous Chondrocyte Implantation

Textor

Hoburg

Lehnigk

et al. 2023

IJMS

Self Cite

View full text Add to dashboard Cite

Loose bodies (LBs) from patients with osteochondritis dissecans (OCD) are usually removed and discarded during surgical treatment of the defect. In this study, we address the question of whether these LBs contain sufficient viable and functional chondrocytes that could serve as a source for autologous chondrocyte implantation (ACI) and how the required prolonged in vitro expansion affects their phenotype. Chondrocytes were isolated from LBs of 18 patients and compared with control chondrocyte from non-weight-bearing joint regions (n = 7) and bone marrow mesenchymal stromal cells (BMSCs, n = 6) obtained during primary arthroplasty. No significant differences in the initial cell yield per isolation and the expression of the chondrocyte progenitor cell markers CD44 + /CD146+ were found between chondrocyte populations from LBs (LB-CH) and control patients (Ctrl-CH). During long-term expansion, LB-CH exhibited comparable viability and proliferation rates to control cells and no ultimate cell cycle arrest was observed within 12 passages respectively 15.3 ± 1.1 mean cumulative populations doublings (CPD). The chondrogenic differentiation potential was comparable between LB-CH and Ctrl-CH, but both groups showed a significantly higher ability to form a hyaline cartilage matrix in vitro than BMSC. Our data suggest that LBs are a promising cell source for obtaining qualitatively and quantitatively suitable chondrocytes for therapeutic applications, thereby circumventing donor site morbidity as a consequence of the biopsies required for the current ACI procedure.

show abstract

Batch Effects during Human Bone Marrow Stromal Cell Propagation Prevail Donor Variation and Culture Duration: Impact on Genotype, Phenotype and Function

Cited by 12 publications

References 69 publications

The enhancer landscape predetermines the skeletal regeneration capacity of stromal cells

The enhancer landscape predetermines the skeletal regeneration capacity of stromal cells

Development of serum substitute medium for bone tissue engineering

Chondrocyte Isolation from Loose Bodies—An Option for Reducing Donor Site Morbidity for Autologous Chondrocyte Implantation

Contact Info

Product

Resources

About