Chondrogenesis from umbilical cord blood cells stimulated with BMP-2 and BMP-6

Abstract

Umbilical cord blood contains undifferentiated mesenchymal stem cells (MSCs) with chondrogenic potential that may be used for the repair of joint damage. The role of growth factors during the process of chondrogenesis is still not entirely understood. The objective of this study was to evaluate the formation of chondrocytes, cartilaginous matrix and type II collagen from human umbilical cord blood stem cells exposed to two different growth factors, BMP-6 and BMP-2, while being cultured as a micromass or a monolayer. Umbilical cord blood was obtained from full-term deliveries, and then, mononuclear cells were separated and cultured for expansion. Afterward, these cells were evaluated by flow cytometry using antibodies specific for MSCs and induced to chondrogenic differentiation in micromass and monolayer cultures supplemented with BMP-2 and BMP-6. Cellular phenotype was evaluated after 7, 14 and 21 days by RT-PCR and Western blot analysis to identify the type II collagen and aggrecan. The expanded cells displayed surface antigens characteristic of mesenchymal progenitor cells and were negative for hematopoietic differentiation antigens. Type II collagen and aggrecan mRNAs were expressed from day 14 in cells stimulated with BMP-2 or BMP-6. Type II collagen was demonstrated by Western blotting in both groups, and the greatest expression was observed 21 days after the cells were stimulated with BMP-2 cultured in micromass. BMP-2 in micromass culture was more efficient to induce the chondrogenesis.

High seeding density of human chondrocytes in agarose produces tissue-engineered cartilage approaching native mechanical and biochemical properties

“…When attempting to grow human cartilage in vitro, human mesenchymal stem cells (MSCs) have commonly served as cell sources (Bhumiratana et al, 2014; Bian et al, 2011; de Mara et al, 2013; Ghone and Grayson, 2012; Handorf and Li, 2011). While MSCs are readily obtained from patients from various tissues, they must be coaxed and maintained along chondrogenic lineage.…”

Human Fetal and Adult Bone Marrow-Derived Mesenchymal Stem Cells Use Different Signaling Pathways for the Initiation of Chondrogenesis

“…Human perinatal fetal stem cells harvested from prenatal and extraembryonic tissues, including fetal bone marrow, blood, liver, amniotic fluid, and umbilical cord blood, have shown trilineage differentiation potential along adipogenic, osteogenic, and chondrogenic lineages [12,[17][18][19][20][21]. However, their potential to generate mature chondrocytes suitable for cartilage tissue engineering has not yet been fully established.…”

Bone and cartilage regeneration with the use of umbilical cord mesenchymal stem cells

“…Compared WJMSCs with temporomandibular chondrocytes in PGA scaffolds and 1 month static cultures WJMSC constructs produced higher amounts of collagen I, more GAGs and had a 55% higher cellularity than chondrocyte constructs Human WJMSCs may have the potential to be used in cartilage engineering although they mainly produce type I collagen [72] de Mara et al Constructs with 4% hyaluronic acid hydrogels created microscopically and macroscopically better cartilage as proved by the levels and organization of collagen type II bundles inside the implanted constructs Human UCBMSCs combined with hyaluronic hydrogels are an attractive option for cartilage regeneration [75] Zheng et al Co-culturing human UCBMSCs with chondrocytes and IGF-1 yields sufficient cartilage ECM production [77] Unless stated otherwise, chondrogenesis was induced with classic chondrogenic medium containing insulin-transferrin-selenium, non-essential amino acids, dexamethasone, L-proline, TGF-b, sodium pyruvate and ascorbic acid, for 21 days. differentiation process compared to BM-MSCs [50,53].…”

“…They phenotypically resemble BM-MSCs and are able to form lacunae-like structures and produce type II collagen [74], which is further increased when stimulated with BMP-6 [75]. Moreover, when seeded on 4% hyaluronic acid hydrogels and implanted into femoral defects they managed to cover them with tissue containing type II collagen, which was macroscopically similar to native cartilage [76].…”