Cryopreserved human adipogenic-differentiated pre-adipocytes: a potential new source for adipose tissue regeneration

Kim, M.H.; Kim, I.; Kim, S.-H.; Jung, M.; Sun, Hongli; Lee, Jangho; Nam, Jinwoo; Lee, S.-K.; Bang, Sa Ik

doi:10.1080/14653240701358452

Cited by 27 publications

(11 citation statements)

References 22 publications

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“…Such combinatorial knockdown would further decrease the stability of the fibroblast network where a chemical cocktail may provoke trans -differentiation into neighboring cell types. Since the protocols for adipogenic differentiation from MSCs and pre-adipocytes have been well established (21–23), we investigated the fibroblast-to-adipocyte transition after suppressing the fibroblastic network followed by repeated inductions of IBMX, dexamethasone, indomethacin and insulin, hereby referred to as adipogenic-induction medium (Figure 2A). …”

Section: Resultsmentioning

confidence: 99%

A transient disruption of fibroblastic transcriptional regulatory network facilitatestrans-differentiation

Tomaru

Hasegawa²,

Suzuki³

et al. 2014

Nucleic Acids Res

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Transcriptional Regulatory Networks (TRNs) coordinate multiple transcription factors (TFs) in concert to maintain tissue homeostasis and cellular function. The re-establishment of target cell TRNs has been previously implicated in direct trans-differentiation studies where the newly introduced TFs switch on a set of key regulatory factors to induce de novo expression and function. However, the extent to which TRNs in starting cell types, such as dermal fibroblasts, protect cells from undergoing cellular reprogramming remains largely unexplored. In order to identify TFs specific to maintaining the fibroblast state, we performed systematic knockdown of 18 fibroblast-enriched TFs and analyzed differential mRNA expression against the same 18 genes, building a Matrix-RNAi. The resulting expression matrix revealed seven highly interconnected TFs. Interestingly, suppressing four out of seven TFs generated lipid droplets and induced PPARG and CEBPA expression in the presence of adipocyte-inducing medium only, while negative control knockdown cells maintained fibroblastic character in the same induction regime. Global gene expression analyses further revealed that the knockdown-induced adipocytes expressed genes associated with lipid metabolism and significantly suppressed fibroblast genes. Overall, this study reveals the critical role of the TRN in protecting cells against aberrant reprogramming, and demonstrates the vulnerability of donor cell's TRNs, offering a novel strategy to induce transgene-free trans-differentiations.

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Section: Resultsmentioning

confidence: 99%

A transient disruption of fibroblastic transcriptional regulatory network facilitatestrans-differentiation

Tomaru

Hasegawa²,

Suzuki³

et al. 2014

Nucleic Acids Res

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“…Complications such as cyst formation or microcalcifications occur in less than 2 to 3% of patients. Clinical trial reports in abstract form conducted by surgeons in Spain, South Korea, and elsewhere suggest similar outcomes [12]. …”

Section: Published Clinical Study Reportsmentioning

confidence: 99%

Clinical and preclinical translation of cell-based therapies using adipose tissue-derived cells

2010

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Adipose tissue is now recognized as an accessible, abundant, and reliable site for the isolation of adult stem cells suitable for tissue engineering and regenerative medicine applications. The past decade has witnessed an explosion of preclinical data relating to the isolation, characterization, cryopreservation, differentiation, and transplantation of freshly isolated stromal vascular fraction cells and adherent, culture-expanded, adipose-derived stromal/stem cells in vitro and in animal models. This body of work has provided evidence supporting clinical translational applications of adipose-derived cells in safety and efficacy trials. The present article reviews the case reports and phase I-III clinical evidence using autologous adipose-derived cells that have been published, to date, in the fields of gastroenterology, neurology, orthopedics, reconstructive surgery, and related clinical disciplines. Future directions and challenges facing the field are discussed and evaluated.

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“…8,34,54,55,59 Fortunately, adipose tissue is an easy and abundant source for such adult stem cells and long-term storage (cryopreservation) procedures for these cells are currently being actively explored. 11,[18][19][20]22,35,[50][51][52][53]62 To alleviate the concerns with the in vivo use of commonly used cryoprotective chemicals (dimethylsulfoxide, glycerol), our laboratory has recently completed a series of studies on ASCs that demonstrate the utility of PVP as a CPA for ASCs. [51][52][53] Briefly, Thirumala et al 53 found that the viability of ASCs frozen in PVP and Dulbecco's Modified Eagle Medium (DMEM) resembles an inverted U-shape (with 10% PVP being optimal).…”

Section: Introductionmentioning

confidence: 99%

Polyvinylpyrrolidone (PVP) Mitigates the Damaging Effects of Intracellular Ice Formation in Adult Stem Cells

Guha

Devireddy

2010

Ann Biomed Eng

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The objective of this work was to assess the effect of 10% (w/v) polyvinylpyrrolidone (PVP) on the pattern of intracellular ice formation (IIF) in human adipose tissue derived adult stem cells (ASCs) in the absence of serum and other cryoprotective agents (CPAs). The freezing experiments were carried out using a fluorescence microscope equipped with a Linkam cooling stage using two cooling protocols. Both the cooling protocols had a common cooling ramp: cells were cooled from 20 degrees C to -8 degrees C at 20 degrees C/min and then further cooled to -13 degrees C at 1 degrees C/min. At this point we employed either cooling protocol 1: the cells were cooled from -13 degrees C to -40 degrees C at a pre-determined cooling rate of 1, 5, 10, 20, or 40 degrees C/min and then thawed back to 20 degrees C at 20 degrees C/min; or cooling protocol 2: the cells were re-warmed from -13 degrees C to -5 degrees C at 20 degrees C/min and then re-cooled at a pre-determined rate of 1, 5, 10, 20, or 40 degrees C/min to -40 degrees C. Almost all (>95%) of the ASCs frozen in 1x PBS and protocol 1 exhibited IIF. However, almost none (<5%) of the ASCs frozen in 1x PBS and protocol 2 exhibited IIF. Similarly, almost all (>95%) of the ASCs frozen in 10% PVP in PBS and protocol 1 exhibited IIF. However, ~0, ~40, ~47, ~67, and ~100% of the ASCs exhibited IIF when frozen in 10% PVP in PBS and utilizing protocol 2 at a cooling rate of 1, 5, 10, 20, or 40 degrees C/min, respectively.

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Cryopreserved human adipogenic-differentiated pre-adipocytes: a potential new source for adipose tissue regeneration

Cited by 27 publications

References 22 publications

A transient disruption of fibroblastic transcriptional regulatory network facilitatestrans-differentiation

A transient disruption of fibroblastic transcriptional regulatory network facilitatestrans-differentiation

Clinical and preclinical translation of cell-based therapies using adipose tissue-derived cells

Polyvinylpyrrolidone (PVP) Mitigates the Damaging Effects of Intracellular Ice Formation in Adult Stem Cells

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