Comparison of the differentiation abilities of bone marrow‑derived mesenchymal stem cells and adipose‑derived mesenchymal stem cells toward nucleus pulposus‑like cells in three‑dimensional culture

Abstract

Nucleus pulposus cell (NPC) transplantation can be a potential therapeutic approach for intervertebral disc degeneration (IDD). However, low cell viability has restricted the therapeutic capacity of NPCs, and sources of natural NPCs are limited. Bone marrow-derived mesenchymal stem cells (BMSCs) and adipose-derived mesenchymal stem cells (ADSCs) can be differentiated toward NPC-like cells. However, it is unknown whether there are differences in the abilities of these two cell types to differentiate into NPC-like cells, or which cell type exhibits the best differentiation ability. The present study compared the abilities of BMSCs and ADSCs to differentiate toward NPC-like cells with or without a 3D culture system to lay a foundation for stem cell transplantation therapy for IDD. BMSCs were isolated from the rat whole bone marrow cell using the repeated adherent culture method. ADSCs were isolated from rat adipose tissues in the subcutaneous inguinal region using the enzyme digestion method. Cells were identified using flow cytometry. Cell viability was assessed via Cell Counting Kit-8 assays, and reverse transcription-quantitative PCR and western blotting were carried out to evaluate the expression of NPC markers and chondrocyte-specific genes. Glycosaminoglycans (GAGs) and proteoglycans were examined via Alcian blue and safranin O staining, respectively. ADSCs in 3D culture displayed the highest cell proliferative ability, compared with the 2D culture system and BMSC culture. In addition, ADSCs in 3D culture exhibited increased GAG and proteoglycan synthesis than BMSCs. Compared with BMSCs in 3D culture, ADSCs in 3D culture exhibited higher mRNA and protein expression of NPC marker genes (hypoxia-inducible factor 1-α, glucose transporter 1) and chondrocyte-specific genes (Sox-9, aggrecan and type II collagen). The present findings indicated that ADSCs exhibited a better ability to differentiate into NPC-like cells in 3D culture compared with BMSCs, which may be of value for the regeneration of intervertebral discs using cell transplantation therapy.

Direct Reprogramming and Induction of Human Dermal Fibroblasts to Differentiate into iPS-Derived Nucleus Pulposus-like Cells in 3D Culture

“…Thus, 3D culture with alginate beads combined with TGF-β signaling is required for directly differentiating and reprogramming NHDFs to generate NP-like cells in vitro. Additionally, considering potential targets of sources for cell therapy as described above ( Figure 7 ), induced pluripotent stem cells [ 23 , 24 ], adipose-derived or bone marrow-derived mesenchymal stem cells [ 42 , 43 , 46 ], stem cells-derived cartilage endplate cells [ 44 ], and adipocytes [ 45 ] have been reported. Here, we found that NHDF differentiates into NP-like cells by inducing defined factors.…”

Application of stem cells in the repair of intervertebral disc degeneration

“…When rat BMSCs and ADMSCs were separately cultured in a three-dimensional culture system and compared with BMSCs in three-dimensional culture, ADMSCs in three-dimensional culture exhibited higher mRNA and protein expression of NP cell marker genes such as HIF1-α, glucose transporter-1, and chondrocytespecific genes such as Sox-9, ACAN, and COL II. These findings indicated that ADMSCs had better ability to differentiate into NP cells in three-dimensional culture [69]. In a study on co-culture of ADMSCs and degenerative NP cells, Han et al [70] found that degenerative NP cells exhibited differential expression of lncRNAs and mRNAs, and subsequently explored the relationship between ADMSCs and NP cells in this co-culture system at the gene expression level.…”

Intervertebral Disc Progenitors: Lessons Learned from Single-Cell RNA Sequencing and the Role in Intervertebral Disc Regeneration

“…Cytometry results revealed that adipose-derived stem cells (ADSCs) were positive for CD73, CD105, CD44 and Sca-1 [ 56 ] and negative for CD34, CD11b and CD45 [ 57 ]. NP-like [ 57 ] and adipocyte differentiation can be induced in ADSCs.…”

“…NP-like [ 57 ] and adipocyte differentiation can be induced in ADSCs. Compared to BMSCs, ADSCs exhibited a higher potential for differentiating into NP-like cells, as shown by both genetic and mRNA analysis [ 56 ]. Staining also revealed the osteogenic and chondrogenic potential of ADSCs [ 58 ].…”

“…Staining also revealed the osteogenic and chondrogenic potential of ADSCs [ 58 ]. In addition, ADSCs were found to proliferate faster than BMSCs in both 3D and 2D cultures [ 56 ]. However, another study found that unstimulated ADSCs and BMSCs had similar proliferation abilities, which may be ascribed to different test methods.…”