Chemically defined serum-free conditions for cartilage regeneration from human embryonic stem cells

Yang, Dandan; Chen, Shubin; Gao, Changzhao; Liu, Xiaobo; Zhou, Yulai; Liu, Pengfei; Cai, Jinglei

doi:10.1016/j.lfs.2016.09.004

Cited by 12 publications

(6 citation statements)

References 35 publications

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“…Moreover, after eight weeks of subcutaneous sphere transplantation in a mouse model, it enabled maturation in non-hypertrophic cartilage. The obtained cells exhibited a strong intensity of a staining characteristic for cartilage [49]. A similar observation was made by Yamashita’s group.…”

Section: Discussionsupporting

confidence: 71%

“…That approach provided a more homogenous population, but the use of animal-derived serum excludes its usefulness in clinical application [51]. In general, these studies emphasize the important fact that a microenvironment, prolonged differentiation in case of chondrogenic lineage is necessary for a derivation suitable product for regenerative purposes and this further supports our claim of obtaining an early chondrocyte-like cell population [49,50,51].…”

Section: Discussionmentioning

confidence: 84%

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Chondrogenic Differentiation of Pluripotent Stem Cells under Controllable Serum-Free Conditions

Lach

Wróblewska

Kulcenty

et al. 2019

IJMS

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The repair of damaged articular cartilage using currently available implantation techniques is not sufficient for the full recovery of patients. Pluripotent stem cells (iPSC)-based therapies could bring new perspectives in the treatment of joint diseases. A number of protocols of in vitro differentiation of iPSC in chondrocytes for regenerative purposes have been recently described. However, in order to use these cells in clinics, the elimination of animal serum and feeder cells is essential. In our study, a strictly defined and controllable protocol was designed for the differentiation of pluripotent stem cells (BG01V, ND 41658*H, GPCCi001-A) in chondrocyte-like cells in serum- and a feeder cell-free system, using the embryoid bodies step. The extension of the protocol and culture conditions (monolayer versus 3D culture) was also tested after the initial 21 days of chondrogenic differentiation. Promotion of the chondrogenic differentiation in 3D culture via the elevated expression of genes related to chondrogenesis was achieved. Using immunofluorescence and immunohistochemistry staining techniques, the increased deposition of the specific extracellular matrix was indicated. As a result, chondrocyte-like cells in the early stages of their differentiation using pellet culture under fully controlled and defined conditions were obtained.

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Section: Discussionsupporting

confidence: 71%

Section: Discussionmentioning

confidence: 84%

Chondrogenic Differentiation of Pluripotent Stem Cells under Controllable Serum-Free Conditions

Lach

Wróblewska

Kulcenty

et al. 2019

IJMS

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“…the subcutaneous transplantation (Yang et al, 2016). Therefore, transplantation to the subcapsular kidney space and the subcutaneous space can both reveal the full developmental potential of transplanted tissue.…”

Section: Discussionmentioning

confidence: 99%

Transmembranous and enchondral osteogenesis in transplants of rat limb buds cultivated in serum‐ and protein‐free culture medium

Himelreich-Perić¹,

Mužić-Radović²,

Marić³

et al. 2022

Anat Histol Embryol

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Cartilage differentiates in rat limb buds cultivated in a chemically defined protein‐free medium in the same manner as in the richer serum‐supplemented medium. We aimed to investigate the remaining differentiation potential of pre‐cultivated limb buds by subsequent transplantation in vivo. Rat front (FLBs) and hind‐limb buds (HLBs) were isolated from Fischer rat dams at the 14th gestation day (GD 14) and cultivated at the air‐liquid interface in Eagle's Minimum Essential Medium (MEM) alone; with 5 μM of 5‐azacytidine (5azaC) or with rat serum (1:1). Overall growth was measured seven times during the culture by an ocular micrometre. After 14 days, explants were transplanted under the kidney capsule of adult males. Growth of limb buds was significantly lower in all limb buds cultivated in MEM than in those cultivated with serum. In MEM with 5azaC, growth of LBs was significantly lower only on day 3 of culture. Afterwards, it was higher throughout the culture period, although a statistically significant difference was assessed only for HLBs. In transplants, mixed structures developed with the differentiated transmembranous bone, cartilage with enchondral ossification, bone‐marrow, sebaceous gland, and hair that have never been found in vitro. Nerves differentiated only in transplants precultivated in the serum‐supplemented medium. We conclude that pre‐cultivation of LBs in a chemically defined protein‐free medium does not restrict osteogenesis and formation of epidermal appendages but is restrictive for neural tissue. These results are important for understanding limb development and regenerative medicine strategies.

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“…However, for their further maturation, the extended time of differentiation and microenvironmental stimulation is necessary. Recently reported in vivo studies of chondrogenic PSC differentiation by several independent groups have shown that implantation of prochondrogenic pellets (around 30 days of 3D culture) with low-deposited proteoglycans enabled formation hyaline cartilage in the damaged site [36,[52][53][54]. One of the studies suggested that early stage for implantation could improve integration with donor tissue because in the prolonged cultured chondrogenic pellets formation of lubricious phase on their surface is observed [36,55].…”

Section: Discussionmentioning

confidence: 99%

The effect of the combination of TGF-β1 and BMP2 with high-density pellet cell culture during chondrogenic differentiation of pluripotent stem cells.

Lach

Suchorska

2020

LOS

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Introduction: The osteoarthritis is a serious threat for well-developed and ageing countries. Present techniques of treatment of damaged cartilage are not sufficient. Hence, new strategies should be developed. One of the potential sources for the regeneration of cartilage is pluripotent stem cells (PSC). Aim: The development of an efficient protocol of chondrogenic differentiation using PSC. Material and methods: The human embryonic stem cell line (BG01V) was used in this study. The chondrogenic differentiation was performed using high-density pellet culture in the presence of TGF-β1 (10 ng/ml) and BMP2 (100 ng/ml). After 21 days gene expression analysis of markers related to chondrogenesis was done. Additionally, the histological staining was performed to detect the deposition of proteoglycans and collagens in differentiated pellet culture. Results: Obtained pellets exhibited decreased expression of pluripotent markers. The upregulation of mesodermal marker and type II collagen was observed in differentiated pellets in the presence of applied growth factors. The histological analysis revealed mild deposition of proteoglycans and collagens. Conclusion: The presented approach enables to obtain chondrogenic pellets in their early stages of chondrogenesis.

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Chemically defined serum-free conditions for cartilage regeneration from human embryonic stem cells

Cited by 12 publications

References 35 publications

Chondrogenic Differentiation of Pluripotent Stem Cells under Controllable Serum-Free Conditions

Chondrogenic Differentiation of Pluripotent Stem Cells under Controllable Serum-Free Conditions

Transmembranous and enchondral osteogenesis in transplants of rat limb buds cultivated in serum‐ and protein‐free culture medium

The effect of the combination of TGF-β1 and BMP2 with high-density pellet cell culture during chondrogenic differentiation of pluripotent stem cells.

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