scholarly journals Feeder & basic fibroblast growth factor-free culture of human embryonic stem cells: Role of conditioned medium from immortalized human feeders

2016 ◽  
Vol 144 (6) ◽  
pp. 838 ◽  
Author(s):  
Sujata Mohanty ◽  
Pooja Teotia ◽  
Shilpa Sharma ◽  
Balram Airan
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Conditioned Medium ◽  
Basic Fibroblast Growth Factor ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Fibroblast Growth

scholarly journals Expression and Potential Role of Fibroblast Growth Factor 2 and Its Receptors in Human Embryonic Stem Cells

Stem Cells ◽  
2005 ◽  
Vol 23 (8) ◽  
pp. 1200-1211 ◽  
Author(s):  
Petr Dvorak ◽  
Dana Dvorakova ◽  
Stanislava Koskova ◽  
Martina Vodinska ◽  
Miroslava Najvirtova ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Human Embryonic Stem Cells ◽  
Potential Role ◽  
Embryonic Stem ◽  
Fibroblast Growth Factor 2 ◽  
Fibroblast Growth

Noggin, Retinoids, and Fibroblast Growth Factor Regulate Hepatic or Pancreatic Fate of Human Embryonic Stem Cells

2010 ◽  
Vol 138 (7) ◽  
pp. 2233-2245.e14 ◽  
Author(s):  
Josué Kunjom Mfopou ◽  
Bing Chen ◽  
Ileana Mateizel ◽  
Karen Sermon ◽  
Luc Bouwens
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Fibroblast Growth

Fibroblast Growth Factor Receptors in Human Embryonic Stem Cells.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4168-4168
Author(s):  
Dana Dvorakova ◽  
Stanislava Koskova ◽  
Martina Vodinska ◽  
Ales Hampl ◽  
Jiri Mayer ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Basic Fibroblast Growth Factor ◽  
Embryonic Stem ◽  
Embryoid Bodies ◽  
Fibroblast Growth ◽  
Fibroblast Growth Factor Receptors ◽  
Aggregation Step

Abstract Signals via fibroblast growth factor receptors (FGFRs) are involved in mesoderm induction events and may be also critical for early hematopoietic specification and proliferation of the hemangioblast. In vitro differentiated embryonic stem cells represent excellent system for the study of early hematopoietic commitment, particularly for understanding signals regulating the onset of hematopoietic differentiation. We have used human embryonic stem cells (hESCs) to study the expression of FGFR1, 2, 3, and 4 in undifferentiated cells and their differentiated progeny. Culturing hESCs i/ in high densities (protocol 1), ii/ without feeder layer of mouse embryonal fibroblasts and basic fibroblast growth factor (protocol 2), and iii/ in three-dimensional aggregates called embryoid bodies (protocol 3), was used to induce the differentiation. To achieve more directed and homogenous differentiation feeder-free hESCs were first subjected to the aggregation step (formation of embryoid bodies) that resembles the gastrulation process. This was followed by differentiation in monolayer in the presence of basic fibroblast growth factor (protocol 4). Such two-step differentiation protocol (5 + 10 days) was shown to activate ectodermal and mesodermal genes and form ectodermal and mesodermal cells (Schuldiner et al., PNAS97:11307, 2000). The gene expression levels for all FGFRs were determined by quantitative real-time RT-PCR. Real-time RT-PCR results were normalized by comparison to the expression of ABL gene. We revealed that undifferentiated hESCs that were cultured with feeder cells and in low density express all four FGFRs in the following pattern: FGFR1 is highly expressed and dominant; FGFR3 is also strongly expressed; FGFR4 shows lower expression; and FGFR2 is only weakly expressed. This expression pattern was changed when hESCs grew and started to differentiate in high densities (protocol 1) or have initiated differentiation either by feeder cells and basic fibroblast growth factor withdrawal or by aggregation step (protocol 2 and 3). Two-fold upregulation of FGFR1 and FGFR4, and downregulation of FGFR3 characterize such changed expression pattern. Notably, the expression levels for all four FGFRs were increased when hESCc were subjected to the two-step differentiation protocol (protocol 4). Compared to the undifferentiated hESCs, FGFR1 and 4 exhibited 7-fold increase, and FGFR2 and 3 were found to be upregulated more than twice. In summary our results show that the expression of FGFRs tightly follows changing culture conditions that may direct hESCs to differentiate. Furthermore, strong upregulation of FGFR1 and 4 in prospective hESC-derived mesodermal cells suggests their involvement in the earliest stages of hematopoiesis. This research was supported in part by the Grant Agency of the Czech Republic (301/03/1122), Ministry of Health (MZ 00065269705), Ministry of Education, Youth, and Sports (MSM 432100001, LN 00A065), and Academy of Sciences of the Czech Republic (AV 0Z5039906).

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